Difference between revisions of "Team:Rotterdam HR/Notebook"

 
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<head>
 
<head>
 
<meta name="viewport" content="width=device-width, initial-scale=1">
 
<meta name="viewport" content="width=device-width, initial-scale=1">
−
<link rel="stylesheet" href="https://2018.igem.org/Team:Rotterdam_HR/css/main?action=raw&ctype=text/css">
+
<link rel="stylesheet" href="https://2018.igem.org/Template:Rotterdam_HR/css/main?action=raw&ctype=text/css">
−
 
+
 
<style>
 
<style>
−
+
 
−
.menu #project, .menu #notebook {
+
.menu #notebook, .menu #project-category {
 
color: goldenrod;
 
color: goldenrod;
 
}
 
}
 
 
−
.cards {
+
</style>
−
display: flex;
+
</head>
−
flex-wrap: wrap;
+
<body>
−
justify-content: center;
+
−
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+
−
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+
−
}
+
  
−
.card {
+
<style>
−
width: 180px;
+
.header::before {
−
height: 180px;
+
background: black url("https://static.igem.org/mediawiki/2018/3/3e/T--Rotterdam_HR--Notebook-banner.jpeg") no-repeat left;
−
margin: 10px;
+
  background-size: cover;
−
position: relative;
+
}
−
text-align: center;
+
−
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+
−
transition: 200ms;
+
−
border-radius: 3px;
+
−
color: white;
+
−
text-shadow: 2px 2px rgba(0, 0, 0, 0.1);
+
−
}
+
  
−
.hardware-card {
+
@media screen and (max-width: 720px) {
−
background: var(--hardware-color);
+
.header::before {
 +
background: black url("https://static.igem.org/mediawiki/2018/7/77/T--Rotterdam_HR--Notebook-banner_small.jpeg") no-repeat left;
 +
background-size: cover;
 
}
 
}
 +
}
 +
</style>
  
−
.wetlab-card {
+
<div class="header">
−
background: var(--wetlab-color);
+
  <div>
−
}
+
<div id="icon"></div>
 +
<script> $('#icon').load("https://2018.igem.org/Template:Rotterdam_HR/icon/notebook?action=raw&ctype=text/html"); </script>
 +
    <h1 style="background: none;">Notebook</h1>
 +
  </div>
 +
</div>
 +
<div class="page page-notebook">
 +
<div id="navbar" style="position: -webkit-sticky;position: sticky;top: -2px; width: 100%; z-index: 100; margin-bottom: 100px;"></div>
 +
<style>
 +
div.notebook-image {
 +
  display: block !important;
 +
  width: 400px !important;
 +
  margin: 10px auto !important;
 +
}
  
−
.card::before, .card::after {
+
div.notebook-image div.image-holder {
−
content: '';
+
border: rgba(255, 255, 255, 0.1) solid 1px;
−
position: absolute;
+
  width: 400px !important;
−
top: 5px;
+
  height: 400px !important;
−
left: 5px;
+
  background-size: contain !important;
−
right: 5px;
+
  background-repeat: no-repeat !important;
−
bottom: 5px;
+
  background-position: center !important;
−
transition: transform 400ms;
+
}
−
z-index: 0;
+
−
}
+
  
−
.card::before {
+
table {
−
border-top: rgba(255,255,255,0.5) solid 1px;
+
margin: auto !important;
−
border-bottom: rgba(255,255,255,0.5) solid 1px;
+
}
−
transform: scale(0, 1);
+
−
}
+
  
−
.card::after {
+
.cards {
−
border-left: rgba(255,255,255,0.5) solid 1px;
+
display: flex;
−
border-right: rgba(255,255,255,0.5) solid 1px;
+
flex-wrap: wrap;
−
transform: scale(1, 0);
+
max-width: 1200px;
−
}
+
margin: 20px auto;
 +
}
  
−
.card .number {
+
.month {
−
position: absolute;
+
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−
top: 0;
+
font-size: 30px;
−
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+
}
−
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+
−
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+
−
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+
−
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+
−
text-align: left;
+
−
}
+
  
−
.card .title {
+
.card {
−
margin: 0;
+
width: 220px;
−
height: 40px;
+
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−
background: rgba(255, 255, 255, 0.1);
+
margin: 10px;
−
color: white;
+
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−
}
+
text-align: center;
 +
box-shadow: 2px 2px 10px rgba(0, 0, 0, 0.25);
 +
border-radius: 3px;
 +
color: white;
 +
text-shadow: 2px 2px rgba(0, 0, 0, 0.1);
 +
display: none;
 +
}
  
−
.card .more-info {
+
.hardware-card {
−
transform: scale(0,0);
+
background: var(--color-hardware);
−
position: absolute;
+
}
−
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+
−
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+
−
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+
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+
−
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+
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+
−
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+
−
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+
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+
−
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+
−
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+
−
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+
−
z-index: 1;
+
−
}
+
  
−
.card .more-info:after {
+
.wetlab-card {
−
content: '';
+
background: var(--color-wetlab);
−
display: block;
+
}
−
width: 70%;
+
.software-card {
−
height: 1px;
+
background: var(--color-software);
−
margin: 0 auto;
+
}
−
background: white;
+
−
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+
−
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+
−
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+
−
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+
  
−
.card svg {
+
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−
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−
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+
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−
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−
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+
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−
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+
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−
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+
right: 5px;
 +
bottom: 5px;
 +
transition: transform 400ms;
 +
z-index: 0;
 +
}
  
−
.date {
+
.card::before {
−
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+
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−
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+
border-bottom: rgba(255, 255, 255, 0.5) solid 1px;
−
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+
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−
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+
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−
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+
−
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+
−
transition: transform 400ms;
+
−
color: white;
+
−
}
+
  
−
.card:hover .date {
+
.card::after {
−
transform: scale(0,0);
+
border-left: rgba(255, 255, 255, 0.5) solid 1px;
−
transition: transform 200ms;
+
border-right: rgba(255, 255, 255, 0.5) solid 1px;
−
}
+
transform: scale(1, 0);
 +
}
  
−
.card:hover::before {
+
.card .entryform {
−
transform: scale(1, 1);
+
display: none;
−
}
+
}
  
−
.card:hover::after {
+
.card .titlebar {
−
transform: scale(1, 1);
+
position: relative;
−
}
+
display: flex;
 +
background: rgba(255, 255, 255, 0.1);
 +
color: white;
 +
align-items: center;
 +
padding-bottom: 5px;
 +
text-shadow: 2px 2px rgba(0, 0, 0, 0.1);
 +
flex-direction: column;
 +
min-height: 50px;
 +
}
  
−
.card:hover .title::after {
+
.card .titlebar svg {
−
transform: scale(1, 0.5);
+
position: relative;
−
}
+
top: 0;
 +
left: 0;
 +
right: 0;
 +
bottom: 0;
 +
}
  
−
.card:hover .more-info {
+
.card .titlebar .title,
−
transform: scale(1, 1);
+
.card .titlebar .date,
−
transition: transform 400ms 200ms;
+
.card .titlebar .back {
−
}
+
margin: 0;
 +
color: white;
 +
}
  
−
.card .more-info:hover {
+
.card .titlebar .title {
−
background: rgba(255, 255, 255, 0.3);
+
background: none;
−
}
+
}
  
−
.card .more-info:hover::after {
+
.card .titlebar .date {
−
transform: scale(1, 1);
+
padding: 5px;
−
}
+
}
  
−
.wetlab-filter:checked ~ .cards .hardware-card {
+
.card .titlebar .back {
−
display: none;
+
font-size: 60px;
−
}
+
position: absolute;
 +
top: 15px;
 +
right: 10px;
 +
}
  
−
.hardware-filter:checked ~ .cards .wetlab-card {
+
.card .attendees {
−
display: none;
+
background: rgba(255, 255, 255, 0.2);
−
}
+
color: white;
−
</style>
+
padding: 2px 2px 2px 10px;
 +
}
  
−
</head>
+
.card .description {
−
<body>
+
padding: 10px;
−
<div id="header"></div>
+
color: white !important;
−
<div id="navbar" style="position: -webkit-sticky;position: sticky;top: 0; width: 100%; z-index: 100;"></div>
+
}
  
−
<!-- ------ CONTENT START ------ -->
+
.card .description p {
 +
padding: 10px;
 +
font-size: medium !important;
 +
text-align: center !important;
 +
color: white !important;
 +
}
  
−
<div class="cards">
+
.card .description p.experiment {
−
<div class="card wetlab-card">
+
border-bottom: white solid 2px;
−
<h3 class="title" style="padding: 5px;">CooA Production</h3>
+
}
−
<svg style="width:80px;height:80px" viewBox="0 0 24 24">
+
 
−
<path fill="currentColor" d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z" />
+
.card .description p.experiment ~ p > span:first-child,
−
</svg>
+
.card .description p.experiment ~ p > strong:first-child {
−
<a class="more-info" href="#">Show more</a>
+
font-size: 24px !important;
−
<h2 class="date">19/07/2018</h2>
+
}
 +
 
 +
.card .description span {
 +
color: white !important;
 +
}
 +
 
 +
.card .description table {
 +
background: rgba(0, 0, 0, 0.2) !important;
 +
}
 +
 
 +
.card .description table td {
 +
color: white;
 +
}
 +
 
 +
.card .number {
 +
position: absolute;
 +
top: 0;
 +
left: 0;
 +
width: 30px;
 +
height: 45px;
 +
padding: 2px;
 +
background: linear-gradient(
 +
to bottom right,
 +
rgba(255, 255, 255, 0.1) 0%,
 +
rgba(0, 0, 0, 0.1) 50%,
 +
rgba(255, 255, 255, 0) 50%
 +
);
 +
text-align: left;
 +
}
 +
 
 +
.card .title {
 +
margin: 0;
 +
background: rgba(255, 255, 255, 0.1);
 +
height: auto;
 +
min-height: 45px;
 +
}
 +
 
 +
.card .more-info {
 +
transform: scale(0, 0);
 +
position: absolute;
 +
bottom: 0;
 +
left: 0;
 +
right: 0;
 +
margin: 10px;
 +
padding: 5px;
 +
border-radius: 2px;
 +
box-shadow: 2px 2px 10px rgba(0, 0, 0, 0.2);
 +
text-transform: uppercase;
 +
text-decoration: none;
 +
color: white;
 +
background: rgba(255, 255, 255, 0.5);
 +
transition: transform 400ms;
 +
z-index: 1;
 +
}
 +
 
 +
.card .titlebar .back:hover, .card .more-info:hover {
 +
cursor: pointer;
 +
}
 +
 
 +
.card .more-info:after {
 +
content: "";
 +
display: block;
 +
width: 70%;
 +
height: 1px;
 +
margin: 0 auto;
 +
background: white;
 +
transform: scale(0, 0);
 +
transform-origin: left;
 +
transition: transform 250ms;
 +
}
 +
 
 +
.card svg {
 +
position: absolute;
 +
top: 80px;
 +
left: 70px;
 +
right: 70px;
 +
bottom: 60px;
 +
width: 80px;
 +
height: 80px;
 +
fill: currentColor;
 +
}
 +
 
 +
.card .cardform .date {
 +
position: absolute;
 +
bottom: -10px;
 +
left: 0;
 +
right: 0;
 +
margin-top: 5px;
 +
transform: scale(1, 1);
 +
transition: transform 400ms;
 +
}
 +
 
 +
.card:hover .cardform .date {
 +
transform: scale(0, 0);
 +
transition: transform 200ms;
 +
}
 +
 
 +
.card:hover::before {
 +
transform: scale(1, 1);
 +
}
 +
 
 +
.card:hover::after {
 +
transform: scale(1, 1);
 +
}
 +
 
 +
.card:hover .title::after {
 +
transform: scale(1, 0.5);
 +
}
 +
 
 +
.card:hover .more-info {
 +
transform: scale(1, 1);
 +
transition: transform 400ms 200ms;
 +
}
 +
 
 +
.card .more-info:hover {
 +
background: rgba(255, 255, 255, 0.3);
 +
}
 +
 
 +
.card .more-info:hover::after {
 +
transform: scale(1, 1);
 +
}
 +
 
 +
.wetlab-filter:checked ~ .cards .wetlab-card { display: block; }
 +
.hardware-filter:checked ~ .cards .hardware-card { display: block; }
 +
.software-filter:checked ~ .cards .software-card { display: block; }
 +
 
 +
.switch {
 +
display: none;
 +
}
 +
 
 +
@media screen and (max-width: 1000px) {
 +
.cards {
 +
justify-content: center;
 +
}
 +
}
 +
 
 +
</style>
 +
<h1 class="title">Notebook</h1>
 +
 
 +
<div class="text-container">
 +
<p>
 +
As a team with only one computer scientist (and one electrical engineer who can program),
 +
we wanted a notebook that automatically updates whenever someone adds something to our lab journal.
 +
Luckily, we gained a sponsor that gave use an electronic lab journal as sponsorship.
 +
With this, we can use their API to download specific sections from our lab journal and display them on or wiki.
 +
But, as a multidisciplinary team we have multiple kinds of notebook entries: Software, Hardware and Wetlab.
 +
Therefore we also used the Google Drive RESTful API for downloading software and hardware entries from google sheets files.
 +
</p>
 +
<a class="nav-button" href="https://2018.igem.org/Team:Rotterdam_HR/Software">More info about the notebook generator</a>
 
</div>
 
</div>
 +
 +
<input type="checkbox" class="wetlab-filter" checked><span>show wetlab entries<br></span></input>
 +
<input type="checkbox" class="hardware-filter" checked><span>show hardware entries<br></span></input>
 +
<input type="checkbox" class="software-filter"><span>show software entries<br></span></input>
 +
<div class="cards">
 +
 +
<style>
 +
#switch-1:checked ~ #card-1 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-1:checked ~ #card-1 .entryform {
 +
display: block;
 +
}
 +
#switch-1:checked ~ #card-1 .cardform,
 +
#switch-1:checked ~ #card-1::before,
 +
#switch-1:checked ~ #card-1::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-1" class="switch"></input>
 +
<div class="card software-card" id="card-1" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Team page</h1>
 +
<h3 class="date">May 10</h3>
 +
<label class="back" for="switch-1">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Created the first prototype for the team page
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Team page</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-1" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 10</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-2:checked ~ #card-2 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-2:checked ~ #card-2 .entryform {
 +
display: block;
 +
}
 +
#switch-2:checked ~ #card-2 .cardform,
 +
#switch-2:checked ~ #card-2::before,
 +
#switch-2:checked ~ #card-2::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-2" class="switch"></input>
 +
<div class="card software-card" id="card-2" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Footer</h1>
 +
<h3 class="date">May 20</h3>
 +
<label class="back" for="switch-2">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Social media created
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Footer</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-2" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 20</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-3:checked ~ #card-3 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-3:checked ~ #card-3 .entryform {
 +
display: block;
 +
}
 +
#switch-3:checked ~ #card-3 .cardform,
 +
#switch-3:checked ~ #card-3::before,
 +
#switch-3:checked ~ #card-3::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-3" class="switch"></input>
 +
<div class="card software-card" id="card-3" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Footer</h1>
 +
<h3 class="date">May 21</h3>
 +
<label class="back" for="switch-3">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Social media buttons same color and fixed iGEM style issues
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Footer</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-3" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 21</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-4:checked ~ #card-4 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-4:checked ~ #card-4 .entryform {
 +
display: block;
 +
}
 +
#switch-4:checked ~ #card-4 .cardform,
 +
#switch-4:checked ~ #card-4::before,
 +
#switch-4:checked ~ #card-4::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-4" class="switch"></input>
 +
<div class="card software-card" id="card-4" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Notebook</h1>
 +
<h3 class="date">May 24</h3>
 +
<label class="back" for="switch-4">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Multiple week summary styles created for the notes
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-4" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 24</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-5:checked ~ #card-5 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-5:checked ~ #card-5 .entryform {
 +
display: block;
 +
}
 +
#switch-5:checked ~ #card-5 .cardform,
 +
#switch-5:checked ~ #card-5::before,
 +
#switch-5:checked ~ #card-5::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-5" class="switch"></input>
 +
<div class="card software-card" id="card-5" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Header</h1>
 +
<h3 class="date">May 27</h3>
 +
<label class="back" for="switch-5">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added the header with mobile compatibility als resized logos and readded them to the header
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Header</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-5" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 27</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-6:checked ~ #card-6 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-6:checked ~ #card-6 .entryform {
 +
display: block;
 +
}
 +
#switch-6:checked ~ #card-6 .cardform,
 +
#switch-6:checked ~ #card-6::before,
 +
#switch-6:checked ~ #card-6::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-6" class="switch"></input>
 +
<div class="card software-card" id="card-6" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Main template</h1>
 +
<h3 class="date">May 28</h3>
 +
<label class="back" for="switch-6">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added template header and footer with 'under construction' content
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Main template</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-6" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 28</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-7:checked ~ #card-7 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-7:checked ~ #card-7 .entryform {
 +
display: block;
 +
}
 +
#switch-7:checked ~ #card-7 .cardform,
 +
#switch-7:checked ~ #card-7::before,
 +
#switch-7:checked ~ #card-7::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-7" class="switch"></input>
 +
<div class="card software-card" id="card-7" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Header</h1>
 +
<h3 class="date">May 28</h3>
 +
<label class="back" for="switch-7">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Small fixes and menu adjustments
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Header</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-7" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 28</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-8:checked ~ #card-8 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-8:checked ~ #card-8 .entryform {
 +
display: block;
 +
}
 +
#switch-8:checked ~ #card-8 .cardform,
 +
#switch-8:checked ~ #card-8::before,
 +
#switch-8:checked ~ #card-8::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-8" class="switch"></input>
 +
<div class="card software-card" id="card-8" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Team</h1>
 +
<h3 class="date">May 29</h3>
 +
<label class="back" for="switch-8">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added a PI list with the secondary PI
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Team</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-8" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">May 29</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-9:checked ~ #card-9 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-9:checked ~ #card-9 .entryform {
 +
display: block;
 +
}
 +
#switch-9:checked ~ #card-9 .cardform,
 +
#switch-9:checked ~ #card-9::before,
 +
#switch-9:checked ~ #card-9::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-9" class="switch"></input>
 +
<div class="card software-card" id="card-9" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Main page</h1>
 +
<h3 class="date">June 7</h3>
 +
<label class="back" for="switch-9">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Designed and added content holders to the index Page
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Main page</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-9" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 7</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-10:checked ~ #card-10 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-10:checked ~ #card-10 .entryform {
 +
display: block;
 +
}
 +
#switch-10:checked ~ #card-10 .cardform,
 +
#switch-10:checked ~ #card-10::before,
 +
#switch-10:checked ~ #card-10::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-10" class="switch"></input>
 +
<div class="card software-card" id="card-10" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Team</h1>
 +
<h3 class="date">June 9</h3>
 +
<label class="back" for="switch-10">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added myself to the teammember Page and did some bugfixing
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Team</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-10" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 9</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-11:checked ~ #card-11 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-11:checked ~ #card-11 .entryform {
 +
display: block;
 +
}
 +
#switch-11:checked ~ #card-11 .cardform,
 +
#switch-11:checked ~ #card-11::before,
 +
#switch-11:checked ~ #card-11::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-11" class="switch"></input>
 +
<div class="card software-card" id="card-11" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Header</h1>
 +
<h3 class="date">June 9</h3>
 +
<label class="back" for="switch-11">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Started working on a better looking menubar
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Header</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-11" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 9</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-12:checked ~ #card-12 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-12:checked ~ #card-12 .entryform {
 +
display: block;
 +
}
 +
#switch-12:checked ~ #card-12 .cardform,
 +
#switch-12:checked ~ #card-12::before,
 +
#switch-12:checked ~ #card-12::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-12" class="switch"></input>
 +
<div class="card software-card" id="card-12" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Header</h1>
 +
<h3 class="date">June 10</h3>
 +
<label class="back" for="switch-12">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added the ability to add sub-menus to items in the menu bar. Integration into the website and added indication for which Page you are on
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Header</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-12" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 10</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-13:checked ~ #card-13 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-13:checked ~ #card-13 .entryform {
 +
display: block;
 +
}
 +
#switch-13:checked ~ #card-13 .cardform,
 +
#switch-13:checked ~ #card-13::before,
 +
#switch-13:checked ~ #card-13::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-13" class="switch"></input>
 +
<div class="card software-card" id="card-13" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Construction pages</h1>
 +
<h3 class="date">June 13</h3>
 +
<label class="back" for="switch-13">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Updated all Pages to the new style with an under construction sign
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Construction pages</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-13" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 13</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-14:checked ~ #card-14 {
 +
width: 100%;
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height: auto;
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}
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#switch-14:checked ~ #card-14 .entryform {
 +
display: block;
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}
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#switch-14:checked ~ #card-14 .cardform,
 +
#switch-14:checked ~ #card-14::before,
 +
#switch-14:checked ~ #card-14::after {
 +
display: none;
 +
}
 +
</style>
 +
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<input type="checkbox" id="switch-14" class="switch"></input>
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<div class="card software-card" id="card-14" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Header</h1>
 +
<h3 class="date">June 14</h3>
 +
<label class="back" for="switch-14">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added a way to recognise Pages under construction using the menubar
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Header</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-14" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 14</h2>
 +
</div>
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 +
</div><style>
 +
#switch-15:checked ~ #card-15 {
 +
width: 100%;
 +
height: auto;
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}
 +
#switch-15:checked ~ #card-15 .entryform {
 +
display: block;
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}
 +
#switch-15:checked ~ #card-15 .cardform,
 +
#switch-15:checked ~ #card-15::before,
 +
#switch-15:checked ~ #card-15::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-15" class="switch"></input>
 +
<div class="card software-card" id="card-15" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Main page</h1>
 +
<h3 class="date">June 14</h3>
 +
<label class="back" for="switch-15">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Updated the content containers so that they are responsive
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Main page</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-15" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 14</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-16:checked ~ #card-16 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-16:checked ~ #card-16 .entryform {
 +
display: block;
 +
}
 +
#switch-16:checked ~ #card-16 .cardform,
 +
#switch-16:checked ~ #card-16::before,
 +
#switch-16:checked ~ #card-16::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-16" class="switch"></input>
 +
<div class="card hardware-card" id="card-16" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Architecture design [TC]</h1>
 +
<h3 class="date">June 18</h3>
 +
<label class="back" for="switch-16">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The first draft of the architecture design for the temperature controller was made.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Architecture design [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-16" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 18</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-17:checked ~ #card-17 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-17:checked ~ #card-17 .entryform {
 +
display: block;
 +
}
 +
#switch-17:checked ~ #card-17 .cardform,
 +
#switch-17:checked ~ #card-17::before,
 +
#switch-17:checked ~ #card-17::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-17" class="switch"></input>
 +
<div class="card software-card" id="card-17" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Human Practices page</h1>
 +
<h3 class="date">June 22</h3>
 +
<label class="back" for="switch-17">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Content added to the human practices page
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Human Practices page</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-17" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 22</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-18:checked ~ #card-18 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-18:checked ~ #card-18 .entryform {
 +
display: block;
 +
}
 +
#switch-18:checked ~ #card-18 .cardform,
 +
#switch-18:checked ~ #card-18::before,
 +
#switch-18:checked ~ #card-18::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-18" class="switch"></input>
 +
<div class="card software-card" id="card-18" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Main page</h1>
 +
<h3 class="date">June 23</h3>
 +
<label class="back" for="switch-18">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Content created and added to the main page
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Main page</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-18" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 23</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-19:checked ~ #card-19 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-19:checked ~ #card-19 .entryform {
 +
display: block;
 +
}
 +
#switch-19:checked ~ #card-19 .cardform,
 +
#switch-19:checked ~ #card-19::before,
 +
#switch-19:checked ~ #card-19::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-19" class="switch"></input>
 +
<div class="card hardware-card" id="card-19" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Architecture design [TC]</h1>
 +
<h3 class="date">June 24</h3>
 +
<label class="back" for="switch-19">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Today a few changes have been made to the architecture design of the temperature controller. The choice was made to use a premade power supply as opposed to a self-made one, because this was considering the time left to finish this device the best choice. A few alterations were made to make the architecture design better readable.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Architecture design [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-19" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 24</h2>
 +
</div>
 +
 +
</div><style>
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#switch-20:checked ~ #card-20 {
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width: 100%;
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height: auto;
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#switch-20:checked ~ #card-20 .entryform {
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#switch-20:checked ~ #card-20 .cardform,
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display: none;
 +
}
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</style>
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<input type="checkbox" id="switch-20" class="switch"></input>
 +
<div class="card hardware-card" id="card-20" onclick="">
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 +
<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">First prototype [TC]</h1>
 +
<h3 class="date">June 25</h3>
 +
<label class="back" for="switch-20">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
A 3d drawing was made for the casing that will hold the temperature controller. The first prototype was printed, but was determent to be inadequate due to the lack of enough space for the cooler block.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">First prototype [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-20" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 25</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-21:checked ~ #card-21 {
 +
width: 100%;
 +
height: auto;
 +
}
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#switch-21:checked ~ #card-21 .entryform {
 +
display: block;
 +
}
 +
#switch-21:checked ~ #card-21 .cardform,
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#switch-21:checked ~ #card-21::before,
 +
#switch-21:checked ~ #card-21::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-21" class="switch"></input>
 +
<div class="card software-card" id="card-21" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Team</h1>
 +
<h3 class="date">June 26</h3>
 +
<label class="back" for="switch-21">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added the remaining instructors to the team
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Team</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-21" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 26</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-22:checked ~ #card-22 {
 +
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 +
height: auto;
 +
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 +
#switch-22:checked ~ #card-22 .entryform {
 +
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 +
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 +
#switch-22:checked ~ #card-22 .cardform,
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#switch-22:checked ~ #card-22::before,
 +
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 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-22" class="switch"></input>
 +
<div class="card hardware-card" id="card-22" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Second prototype [TC]</h1>
 +
<h3 class="date">June 26</h3>
 +
<label class="back" for="switch-22">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
In the drawing space was added to fit the cooler block for the temperature controller. Slits were added to the drawing for the nuts to slide into. The altered drawing was printed. The newly printed parts, cooler block, fan and Peltier element were assembled. Everything fit together.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Second prototype [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-22" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 26</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-23:checked ~ #card-23 {
 +
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 +
#switch-23:checked ~ #card-23 .cardform,
 +
#switch-23:checked ~ #card-23::before,
 +
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display: none;
 +
}
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</style>
 +
 +
<input type="checkbox" id="switch-23" class="switch"></input>
 +
<div class="card wetlab-card" id="card-23" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">June 26</h3>
 +
<label class="back" for="switch-23">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
To start testing the production of CooA, we firstly resuspended and transformed the BioBricks from the kit witch we wanted to use (Table 1). This has been done using the iGEM Kit plate instructions (Protocol 1, step 1 - 3) and the NEB 10 b&eacute;ta transformation protocol adapted New England Biolabs (Protocol 2), using bought cells with a reported transformation efficienty of 1-3 10^9 cfu/&mu;g pUC19. Inucbation was at 37&deg;C.<br>
 +
<br>
 +
Table 1 : Resuspended BioBricks from the iGEM 2018 Distribution kit and their uses.
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_K592009</td>
 +
<td style="text-align: center; width: 365px;">Blue Chromoprotein AmilCP</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_B0031</td>
 +
<td style="text-align: center; width: 365px;">Ribosomal binding site (strong), derived from&nbsp;BBa_B0030</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_B0032</td>
 +
<td style="text-align: center; width: 365px;">Ribosomal binding site (medium), derived from&nbsp;BBa_B0030</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_B0030</td>
 +
<td style="text-align: center; width: 365px;">Ribosomal binding site (weak)</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_B0015</td>
 +
<td style="text-align: center; width: 365px;">Double terminator (BBa_B0010 &amp; BBa_B0012)</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_J23100</td>
 +
<td style="text-align: center; width: 365px;">Strong Constitutive Anderson Promotor</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_J23105</td>
 +
<td style="text-align: center; width: 365px;">Medium Constitutive Anderson Promotor</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_J23113</td>
 +
<td style="text-align: center; width: 365px;">Weak Constitutive Anderson Promotor</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_J45199</td>
 +
<td style="text-align: center; width: 365px;">Banana odor enzyme (ATF1) generator</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_K1184000</td>
 +
<td style="text-align: center; width: 365px;">Killer Red</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_K352001</td>
 +
<td style="text-align: center; width: 365px;">CooA</td>
 +
</tr>
 +
<tr>
 +
<td style="text-align: center; width: 122px;">BBa_K352011</td>
 +
<td style="text-align: center; width: 365px;">CooA responsive system</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<br>
 +
After checking the plates on the 28th of June, we recieved the following results (Table 2):<br>
 +
<br>
 +
Table 2 : Kolonies found after overnight growth.
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_K592009</td>
 +
<td style="width: 277px; text-align: center;">20 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_B0031</td>
 +
<td style="width: 277px; text-align: center;">4 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_B0032</td>
 +
<td style="width: 277px; text-align: center;">5 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_B0030</td>
 +
<td style="width: 277px; text-align: center;">1 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_B0015</td>
 +
<td style="width: 277px; text-align: center;">6 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_J23100</td>
 +
<td style="width: 277px; text-align: center;">1 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_J23105</td>
 +
<td style="width: 277px; text-align: center;">98 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_J23113</td>
 +
<td style="width: 277px; text-align: center;">19 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_J45199</td>
 +
<td style="width: 277px; text-align: center;">0 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_K1184000</td>
 +
<td style="width: 277px; text-align: center;">4 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_K352001</td>
 +
<td style="width: 277px; text-align: center;">2 colonies</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 210px; text-align: center;">BBa_K352011</td>
 +
<td style="width: 277px; text-align: center;">4 colonies</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-23" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 26</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-24:checked ~ #card-24 {
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width: 100%;
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height: auto;
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 +
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<input type="checkbox" id="switch-24" class="switch"></input>
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<div class="card hardware-card" id="card-24" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Testrun [TC]</h1>
 +
<h3 class="date">June 27</h3>
 +
<label class="back" for="switch-24">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
A short test run was done with the newly assembled cooler. The test concluded that the temperature controller could cool a water droplet to the point of freezing and heat it back up to its boiling point.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testrun [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-24" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 27</h2>
 +
</div>
 +
 +
</div><style>
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</style>
 +
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<input type="checkbox" id="switch-25" class="switch"></input>
 +
<div class="card wetlab-card" id="card-25" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">June 29</h3>
 +
<label class="back" for="switch-25">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>After making a 20 mL overnight culture of our NEB 10 b&eacute;ta cells containing our BioBricks, we performed a mini prep (Protocol 3). Using our nanodrop spectrophotometer&nbsp;&nbsp;which gave us the following DNA concentrations:<br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td>BBa_K592009</td>
 +
<td>72,5 ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_B0031</td>
 +
<td>118,9&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_B0032</td>
 +
<td>52,5&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_B0030</td>
 +
<td>488,6&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_B0015</td>
 +
<td>53,5&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_J23100</td>
 +
<td>148,3&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_J23105</td>
 +
<td>96,3&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_J23113</td>
 +
<td>151,8&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_K118400</td>
 +
<td>140,4&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_K352001</td>
 +
<td>77,7&nbsp;ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BBa_K352011</td>
 +
<td>76,8&nbsp;ng/&mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<br>
 +
After assessign our results, in the future we will perform the ethanol carry-over steps.<br>
 +
<br>
 +
Also from the previously mentioned overnight culture 1 mL has been used to make Glycerol Stock (Protocol 4)
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-25" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">June 29</h2>
 +
</div>
 +
 +
</div><style>
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#switch-26:checked ~ #card-26 {
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}
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#switch-26:checked ~ #card-26 .cardform,
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#switch-26:checked ~ #card-26::before,
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#switch-26:checked ~ #card-26::after {
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display: none;
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}
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</style>
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<input type="checkbox" id="switch-26" class="switch"></input>
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<div class="card hardware-card" id="card-26" onclick="">
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<div class="entryform">
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<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Draft decision</h1>
 +
<h3 class="date">July 2</h3>
 +
<label class="back" for="switch-26">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The choice to use a computer PSU as a power supply was made. The choice was made on the grounds, that computer PSU have a standard 12, 5 and 3.3V output. This is beneficial because these are the voltages necessary for the selected components.
 +
<p class="experiment"></p>
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</div>
 +
</div>
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 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Draft decision</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-26" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 2</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-27:checked ~ #card-27 {
 +
width: 100%;
 +
height: auto;
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}
 +
#switch-27:checked ~ #card-27 .entryform {
 +
display: block;
 +
}
 +
#switch-27:checked ~ #card-27 .cardform,
 +
#switch-27:checked ~ #card-27::before,
 +
#switch-27:checked ~ #card-27::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-27" class="switch"></input>
 +
<div class="card wetlab-card" id="card-27" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">July 2</h3>
 +
<label class="back" for="switch-27">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>Today we performed a mini prep, digestion, dephosphorilation and ligation (following protocols 4 and 5) of a few promotor and RBS BioBricks. Because of administrative complications only the variants with J23113 and a RBS have been digested correctly.</p>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-27" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 2</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-28:checked ~ #card-28 {
 +
width: 100%;
 +
height: auto;
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}
 +
#switch-28:checked ~ #card-28 .entryform {
 +
display: block;
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}
 +
#switch-28:checked ~ #card-28 .cardform,
 +
#switch-28:checked ~ #card-28::before,
 +
#switch-28:checked ~ #card-28::after {
 +
display: none;
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}
 +
</style>
 +
 +
<input type="checkbox" id="switch-28" class="switch"></input>
 +
<div class="card wetlab-card" id="card-28" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">July 2</h3>
 +
<label class="back" for="switch-28">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>Today we transformed the previously made three combinations (BBa_J23113 + BBa_B0031 / B0032 / B0030) into chemically competent NEb 10 b&eacute;ta E.coli cells using the corresponding NEB protocol.</p>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-28" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 2</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-29:checked ~ #card-29 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-29:checked ~ #card-29 .entryform {
 +
display: block;
 +
}
 +
#switch-29:checked ~ #card-29 .cardform,
 +
#switch-29:checked ~ #card-29::before,
 +
#switch-29:checked ~ #card-29::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-29" class="switch"></input>
 +
<div class="card hardware-card" id="card-29" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
 +
<h1 class="title">Architecture design [TC]</h1>
 +
<h3 class="date">July 5</h3>
 +
<label class="back" for="switch-29">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Changes were made to the architecture design of the temperature controller. Different voltage regulators were selected to fit the newly chosen power supply. To better readability of the architecture design a legend was added.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Architecture design [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-29" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 5</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-30:checked ~ #card-30 {
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width: 100%;
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height: auto;
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display: block;
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}
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#switch-30:checked ~ #card-30 .cardform,
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#switch-30:checked ~ #card-30::before,
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 +
display: none;
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}
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</style>
 +
 +
<input type="checkbox" id="switch-30" class="switch"></input>
 +
<div class="card wetlab-card" id="card-30" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">July 5</h3>
 +
<label class="back" for="switch-30">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>After performing a mini prep and NotI digest on the 5th of July, and a gel electrophoresis today, we concluded that the cloning wasn't succesful.<br>
 +
&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-30" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 5</h2>
 +
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 +
 +
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 +
#switch-31:checked ~ #card-31 {
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width: 100%;
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height: auto;
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#switch-31:checked ~ #card-31 .entryform {
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display: block;
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#switch-31:checked ~ #card-31 .cardform,
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#switch-31:checked ~ #card-31::before,
 +
#switch-31:checked ~ #card-31::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-31" class="switch"></input>
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<div class="card hardware-card" id="card-31" onclick="">
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<div class="entryform">
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<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Electrical circuit design [TC]</h1>
 +
<h3 class="date">July 8</h3>
 +
<label class="back" for="switch-31">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Custom footprints were made for multiple components and added to the parts library in the PCB design software.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Electrical circuit design [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-31" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 8</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-32:checked ~ #card-32 {
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height: auto;
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display: block;
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#switch-32:checked ~ #card-32 .cardform,
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display: none;
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}
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</style>
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<input type="checkbox" id="switch-32" class="switch"></input>
 +
<div class="card hardware-card" id="card-32" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Electrical circuit design [TC]</h1>
 +
<h3 class="date">July 9</h3>
 +
<label class="back" for="switch-32">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The provisional electronic circuit design for the temperature controller was made.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Electrical circuit design [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-32" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 9</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-33:checked ~ #card-33 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-33:checked ~ #card-33 .entryform {
 +
display: block;
 +
}
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#switch-33:checked ~ #card-33 .cardform,
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#switch-33:checked ~ #card-33::after {
 +
display: none;
 +
}
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</style>
 +
 +
<input type="checkbox" id="switch-33" class="switch"></input>
 +
<div class="card wetlab-card" id="card-33" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">July 10</h3>
 +
<label class="back" for="switch-33">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>Today we again performed a mini prep (overnight cultures from 10th of July) and digest. Sadly most digests did not give the expected results. Even though we had already progressed with the ligation. For now we've decided to progress with these ligation products.</p>
 +
 +
<p>&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>06</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-33" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 10</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-34:checked ~ #card-34 {
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#switch-34:checked ~ #card-34 .entryform {
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display: block;
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#switch-34:checked ~ #card-34 .cardform,
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#switch-34:checked ~ #card-34::before,
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#switch-34:checked ~ #card-34::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-34" class="switch"></input>
 +
<div class="card hardware-card" id="card-34" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Electrical circuit design [TC]</h1>
 +
<h3 class="date">July 11</h3>
 +
<label class="back" for="switch-34">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The electronic circuit design for the temperature controller was finished. Measuring point were added to the electronic circuit design.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Electrical circuit design [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-34" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 11</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-35:checked ~ #card-35 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-35:checked ~ #card-35 .entryform {
 +
display: block;
 +
}
 +
#switch-35:checked ~ #card-35 .cardform,
 +
#switch-35:checked ~ #card-35::before,
 +
#switch-35:checked ~ #card-35::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-35" class="switch"></input>
 +
<div class="card wetlab-card" id="card-35" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Stock up</h1>
 +
<h3 class="date">July 11</h3>
 +
<label class="back" for="switch-35">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Loraine Nelson | Paul Reusink</div>
 +
<div class="description">
 +
To have some basic biobricks ready, we stocked them by doing a plasmid isolation and freeze the DNA.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Stock up</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-35" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 11</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-36:checked ~ #card-36 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-36:checked ~ #card-36 .entryform {
 +
display: block;
 +
}
 +
#switch-36:checked ~ #card-36 .cardform,
 +
#switch-36:checked ~ #card-36::before,
 +
#switch-36:checked ~ #card-36::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-36" class="switch"></input>
 +
<div class="card wetlab-card" id="card-36" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Basic parts</h1>
 +
<h3 class="date">July 11</h3>
 +
<label class="back" for="switch-36">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Loraine Nelson | Paul Reusink</div>
 +
<div class="description">
 +
To start on a basic construct we did a digestion and ligation with the parts we isolated earlier. To prevent original constructs we did a defosphorylation after the digestion. We controlled the digestion by doing a gelelktroforeses.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Basic parts</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-36" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 11</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-37:checked ~ #card-37 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-37:checked ~ #card-37 .entryform {
 +
display: block;
 +
}
 +
#switch-37:checked ~ #card-37 .cardform,
 +
#switch-37:checked ~ #card-37::before,
 +
#switch-37:checked ~ #card-37::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-37" class="switch"></input>
 +
<div class="card hardware-card" id="card-37" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Board layout [TC]</h1>
 +
<h3 class="date">July 12</h3>
 +
<label class="back" for="switch-37">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Started working on the board layout for the PCB of the temperature controller.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Board layout [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-37" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 12</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-38:checked ~ #card-38 {
 +
width: 100%;
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height: auto;
 +
}
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#switch-38:checked ~ #card-38 .entryform {
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display: block;
 +
}
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#switch-38:checked ~ #card-38 .cardform,
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#switch-38:checked ~ #card-38::before,
 +
#switch-38:checked ~ #card-38::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-38" class="switch"></input>
 +
<div class="card wetlab-card" id="card-38" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Basic parts</h1>
 +
<h3 class="date">July 12</h3>
 +
<label class="back" for="switch-38">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Randall de Waard</div>
 +
<div class="description">
 +
We continued with the ligated DNA by transformating it to Neb10Beta. This was plated onto agarplates and incubated for 1 day.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Basic parts</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-38" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 12</h2>
 +
</div>
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 +
</div><style>
 +
#switch-39:checked ~ #card-39 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-39:checked ~ #card-39 .entryform {
 +
display: block;
 +
}
 +
#switch-39:checked ~ #card-39 .cardform,
 +
#switch-39:checked ~ #card-39::before,
 +
#switch-39:checked ~ #card-39::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-39" class="switch"></input>
 +
<div class="card hardware-card" id="card-39" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Board layout [TC]</h1>
 +
<h3 class="date">July 13</h3>
 +
<label class="back" for="switch-39">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Further work was done to the board layout for the temperature controller.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Board layout [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-39" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 13</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-40:checked ~ #card-40 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-40:checked ~ #card-40 .entryform {
 +
display: block;
 +
}
 +
#switch-40:checked ~ #card-40 .cardform,
 +
#switch-40:checked ~ #card-40::before,
 +
#switch-40:checked ~ #card-40::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-40" class="switch"></input>
 +
<div class="card wetlab-card" id="card-40" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Basic parts</h1>
 +
<h3 class="date">July 13</h3>
 +
<label class="back" for="switch-40">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
The results of 7/12/2018 were collected, in action of this we decided to plate some ligated and transferred DNA from 7/12/2018 again. This we did because there wasn't a clear grow of colonies on the plates.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Basic parts</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-40" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 13</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-41:checked ~ #card-41 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-41:checked ~ #card-41 .entryform {
 +
display: block;
 +
}
 +
#switch-41:checked ~ #card-41 .cardform,
 +
#switch-41:checked ~ #card-41::before,
 +
#switch-41:checked ~ #card-41::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-41" class="switch"></input>
 +
<div class="card wetlab-card" id="card-41" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Blue White screening on paper</h1>
 +
<h3 class="date">July 13</h3>
 +
<label class="back" for="switch-41">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
We tried to let bacteria grow on paper, therefore we used our own buisnesscards. We let the buisnesscards absorb some LB-medium and put them inside petridishes.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Blue White screening on paper</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-41" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 13</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-42:checked ~ #card-42 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-42:checked ~ #card-42 .entryform {
 +
display: block;
 +
}
 +
#switch-42:checked ~ #card-42 .cardform,
 +
#switch-42:checked ~ #card-42::before,
 +
#switch-42:checked ~ #card-42::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-42" class="switch"></input>
 +
<div class="card hardware-card" id="card-42" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Board layout [TC]</h1>
 +
<h3 class="date">July 15</h3>
 +
<label class="back" for="switch-42">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Work has been done on the board layout for the temperature controller. The board layout is now finished.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Board layout [TC]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">July 15</h2>
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h1 class="title">Assessing CooA Production</h1>
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<h3 class="date">July 15</h3>
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<label class="back" for="switch-43">&times;</label>
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</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
After we transformed our ligation results from the 11th on the 12th, we recieved the following results on the 13th:<br>
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<meta charset="utf-8">
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<p dir="ltr"><br>
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B0030+J23100: No growth<br>
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B0030+J23105: Unsure (small sports, maybe colonies)<br>
 +
B0031+J23100:&nbsp;Growth<br>
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B0031+J23105:&nbsp;Unsure (small sports, maybe colonies)<br>
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B0032+J23100:&nbsp;Growth<br>
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B0032+J23105:&nbsp;Unsure (small sports, maybe colonies)<br>
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K352001+B0015: Growth<br>
 +
<br>
 +
On the 13th we retransformed the 4 ligations of which we didn't get conclusive results, which we looked at today:<br>
 +
B0030+J23100:&nbsp;No growth</p>
 +
 +
<p dir="ltr"><meta charset="utf-8">B0030+J23105: Again unsure<br>
 +
B0031+J23105:&nbsp;No growth<br>
 +
B0032+J23105:&nbsp;No growth<br>
 +
<br>
 +
After assessing the results we made overnight cultures of the following:</p>
 +
 +
<p>B0031+J23100</p>
 +
 +
<p>K352001+B0015</p>
 +
 +
<p>B0032+J23100</p>
 +
 +
<p>B0030+J23105 (1)</p>
 +
 +
<p>B0030+J23105 (2)</p>
 +
 +
<p>B0032+J23105<br>
 +
<br>
 +
Also we retransformed the colonies we retransformed on the 13th again, and quickly spun our cells in a centrifuge, removing most of the supernatant and resuspending our cells before we spread them on Chloramphenicol plates.</p>
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>07</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 15</h2>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Basic parts</h1>
 +
<h3 class="date">July 16</h3>
 +
<label class="back" for="switch-44">&times;</label>
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</div>
 +
<div class="attendees">Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Randall de Waard</div>
 +
<div class="description">
 +
To know if the ligation was succesfull we will controll the DNA, therefor we ented the bacteria's with DNA in LB-medium to grow overnight. We also plated the used bacteria's onto new plates to use for further experiments.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Basic parts</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 16</h2>
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Blue White screening on paper</h1>
 +
<h3 class="date">July 16</h3>
 +
<label class="back" for="switch-45">&times;</label>
 +
</div>
 +
<div class="attendees">Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Randall de Waard</div>
 +
<div class="description">
 +
We did a blue white screening on our buisnesscards to see if bacteria had grew on them. We transferred the buisnesscards to new petrydishes and added ITPG and X-gal to the petrydishes.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Blue White screening on paper</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 16</h2>
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<svg viewBox="0 0 24 24">
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<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<h1 class="title">Notebook</h1>
 +
<h3 class="date">July 17</h3>
 +
<label class="back" for="switch-46">&times;</label>
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</div>
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<div class="attendees">Rens Boeser</div>
 +
<div class="description">
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Frontend design for the notebook
 +
<p class="experiment"></p>
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</div>
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</div>
 +
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook</h3>
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<svg viewBox="0 0 24 24">
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<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<h2 class="date" style="color: white;">July 17</h2>
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<div class="entryform">
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<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Basic parts</h1>
 +
<h3 class="date">July 17</h3>
 +
<label class="back" for="switch-47">&times;</label>
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</div>
 +
<div class="attendees">Rens Boeser | Dustin van der Meulen | Loraine Nelson  | Suzanne Romeijn | Randall de Waard</div>
 +
<div class="description">
 +
The resulst of 7/16/2018 showed that the DNA didnt ligate as planned.  To be able to digest enough DNA we did a plasmid isolation again. We digested this DNA and froze it for further use.
 +
<p class="experiment"></p>
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 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Basic parts</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 17</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Stock up</h1>
 +
<h3 class="date">July 18</h3>
 +
<label class="back" for="switch-48">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Mike Vrieswijk | Randall de Waard</div>
 +
<div class="description">
 +
We did another round of plasmid isolation for stocking.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Stock up</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 18</h2>
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</svg>
 +
<h1 class="title">Notebook</h1>
 +
<h3 class="date">July 19</h3>
 +
<label class="back" for="switch-49">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Designed a way to follow experiments that take up multiple days and eventually designed a notebook entry page and made it a responsive page
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<h2 class="date" style="color: white;">July 19</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Basic parts</h1>
 +
<h3 class="date">July 19</h3>
 +
<label class="back" for="switch-50">&times;</label>
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</div>
 +
<div class="attendees">Dustin van der Meulen | Mei Ju Goemans| Randall de Waard </div>
 +
<div class="description">
 +
We digested all DNA we have in stock to test for our basic construct. We tested the DNA by doing a gelelktroforeses.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>06</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Basic parts</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 19</h2>
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<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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</svg>
 +
<h1 class="title">Notebook</h1>
 +
<h3 class="date">July 21</h3>
 +
<label class="back" for="switch-51">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Created a one page notebook. Made the notebook entry page more suited for the entry length
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<h2 class="date" style="color: white;">July 21</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">July 23</h3>
 +
<label class="back" for="switch-52">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Transformations of the&nbsp;biobricks K133071, K173003 and I13453 are performed. K133071 will produce CO2 if there's pyruvate present and K173003 will produce CO2 if urea is present. I13453 is a promotor which will work if there's arabinose present.&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;<br>
 +
The biobricks were transformed first into electrocometent cells and later&nbsp;into chemical&nbsp;competent cells from the strain NEb10Beta. After transformations the culture was plated on agar plates with&nbsp;antibiotics. The first transformations didn't work out, but the second did, because then the right competent cells were used.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 14px;"><strong>Experiment 1: Transformation biobricks</strong></span><br>
 +
<br>
 +
Transformation of the biobricks K133071, K173003 and I13453 in NEB10B&egrave;ta.<br>
 +
<br>
 +
<span style="font-size:12px;"><strong>Materials&nbsp;</strong></span></p>
 +
 +
<p>- Biobricks (Bba_K133071,&nbsp;Bba_K173003 and Bba_I3453)&nbsp;<br>
 +
- Nuclease free water<br>
 +
- LB- agar<br>
 +
- Variable Volume Pipettes&nbsp;<br>
 +
- Sterile pipetpoints<br>
 +
- Chemically competent cells ( NEB10Beta, 1x10^9)&nbsp;<br>
 +
- Heater (42&deg;C) &nbsp;<br>
 +
- Incubator + shaker (37&deg;C and 250 rpm)&nbsp;<br>
 +
- Eppendorf tubes (1,5 and 2 ml)&nbsp;<br>
 +
- Stable outgrow medium for NEB10Beta or SOC- outgrow medium&nbsp;<br>
 +
-Chloramphenicol (40 mg/mL)<br>
 +
<br>
 +
<span style="font-size:12px;"><strong><span style="font-family:arial,helvetica,sans-serif;">Methods</span></strong></span><br>
 +
-K173003 --&gt; pSB1C3 backbone, plate 6, well 15N<br>
 +
-K133071 --&gt; pSB1C3 backbone, plate 6, wel 21A<br>
 +
-I13453 --&gt; pSB1C3 backbone, plate 3, well 19O</p>
 +
 +
<p>To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24&deg;C). After incubation the DNA is transferred to Eppendorf tubes&nbsp;and put on ice.&nbsp;<br>
 +
<br>
 +
For the transformation 1 ul of the DNA was added&nbsp;to 50 ul&nbsp;chemically competent cells strain&nbsp;NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube&nbsp;in a heater (42&deg;C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes.&nbsp;<br>
 +
For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37&deg;C and 250 rpm.&nbsp;&nbsp;<br>
 +
<br>
 +
After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37&deg;C.&nbsp;<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
The first transformation was done in electrocompetent cells instead of chemocompetent cells so the first transformation did not succeed.</p>
 +
 +
<p>&nbsp;</p>
 +
 +
<p><strong>Conclusion</strong><br>
 +
<br>
 +
The second transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4&deg;C and can be used for further experiments.&nbsp;<br>
 +
The CFU from the&nbsp;negative controle will be plated on a plate with antibiotics to make sure there isn't a plasmid in the bacteria. The outgrow medium will be sterilized by filter&nbsp;again.&nbsp;<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 23</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h1 class="title">ATP sensor</h1>
 +
<h3 class="date">July 24</h3>
 +
<label class="back" for="switch-53">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser | Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Randall de Waard</div>
 +
<div class="description">
 +
New biobricks, used to make an ATP sensor, were transformed into NEB10Beta. The culture was than plated onto agar and incubated for 1 day. 
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">ATP sensor</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 24</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Gas output</h1>
 +
<h3 class="date">July 24</h3>
 +
<label class="back" for="switch-54">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser | Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Randall de Waard</div>
 +
<div class="description">
 +
New biobricks, which produces gasses, were transformed into NEB10Beta. The culture was than plated onto agar and incubated for 1 day. 
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Gas output</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">July 24</h2>
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 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Notebook generator</h1>
 +
<h3 class="date">July 24</h3>
 +
<label class="back" for="switch-55">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Started working on an automatic notebook creator
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook generator</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-55" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 24</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-56:checked ~ #card-56 {
 +
width: 100%;
 +
height: auto;
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}
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#switch-56:checked ~ #card-56 .entryform {
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#switch-56:checked ~ #card-56 .cardform,
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<input type="checkbox" id="switch-56" class="switch"></input>
 +
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 +
 +
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 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">July 24</h3>
 +
<label class="back" for="switch-56">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>To be able to always get the necessary biobricks, there have been&nbsp;made&nbsp;glycerolstocks of the transformed biobricks K133071, K173003 and I13453. The glycerolstocks are stored at -80 degrees Celsius. If needed, they can be retrieved from this storage to use for experiments.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:12px;"><strong>Experiment 2: Glycerolstocks</strong></span><br>
 +
<br>
 +
Making glycerol stocks of the biobricks K133071, K173003, I13453.&nbsp;<br>
 +
<br>
 +
<span style="font-size:12px;"><strong>Materials</strong></span><br>
 +
- 87% glycerol&nbsp;<br>
 +
- culture with biobrick plasmids<br>
 +
- LB-medium<br>
 +
- Variable Volume Pipette<br>
 +
- Sterile pipetpoints<br>
 +
<br>
 +
<strong><span style="font-size:12px;">Methods</span></strong><br>
 +
The colonies from the transformation were used to make an overnight culture. The culture was made by putting a colony in LB-medium and then the culture was&nbsp;incubated for a night at 37&deg;C. &nbsp;<br>
 +
<br>
 +
The next day the cultures had grown close and were ready for a glycerol stock. For making glycerol stocks there was added 250 ul 87% glycerol to 1 ml culture. This was frozen in -80&deg;C. &nbsp;<br>
 +
<br>
 +
The stocks were stored at -80&deg;C in drawer 3, tower 1, drawer&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-56" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 24</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-57:checked ~ #card-57 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-57:checked ~ #card-57 .entryform {
 +
display: block;
 +
}
 +
#switch-57:checked ~ #card-57 .cardform,
 +
#switch-57:checked ~ #card-57::before,
 +
#switch-57:checked ~ #card-57::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-57" class="switch"></input>
 +
<div class="card wetlab-card" id="card-57" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing different ATP sensors</h1>
 +
<h3 class="date">July 24</h3>
 +
<label class="back" for="switch-57">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>Today we did a transformation with&nbsp;new biobricks. The biobricks were built into chemically competent cells from the strain NEb10Beta. After transformation the culture was plated on agar plates with&nbsp;antibiotics. The results were checked the next day.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
Materials:&nbsp;
 +
<p>- Biobricks (Bba-K1499004 and Bba- K1614019)&nbsp;<br>
 +
- Nuclease free water<br>
 +
- LB- agar<br>
 +
- Variable Volume Pipettes&nbsp;<br>
 +
- Sterile pipetpoints<br>
 +
- Chemically competent cells ( NEB10Beta, 1x10^9)&nbsp;<br>
 +
- Heater (42&deg;C) &nbsp;<br>
 +
- Incubator + shaker (37&deg;C and 250 rpm)&nbsp;<br>
 +
- Eppendorf tubes (1,5 and 2 ml)&nbsp;<br>
 +
- Stable outgrow medium for NEB10Beta or SOC- outgrow medium&nbsp;<br>
 +
<br>
 +
Method:&nbsp;<br>
 +
<br>
 +
To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24&deg;C). After incubation the DNA is transferred to Eppendorf tubes&nbsp;and put on ice.&nbsp;<br>
 +
<br>
 +
For the transformation 1 ul of the DNA was added&nbsp;to 50 ul&nbsp;chemically competent cells strain&nbsp;NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube&nbsp;in a heater (42&deg;C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes.&nbsp;<br>
 +
For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37&deg;C and 250 rpm.&nbsp;&nbsp;<br>
 +
<br>
 +
After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37&deg;C.&nbsp;<br>
 +
<br>
 +
Results:<br>
 +
Tabel 1 shows the results of the plates which were incubated.&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 427px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 107px;">Biobricks:&nbsp;</td>
 +
<td style="width: 44px;">Antibiotics:&nbsp;</td>
 +
<td style="width: 86px;">Dilution:&nbsp;</td>
 +
<td style="width: 73px;">Colonies:</td>
 +
<td style="width: 101px;">CFU:&nbsp;</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 107px;">Bba-K1614019</td>
 +
<td style="width: 44px;">&nbsp;Can&nbsp;</td>
 +
<td style="width: 86px;">0x&nbsp;</td>
 +
<td style="width: 73px;">109</td>
 +
<td style="width: 101px;">1*10^3 /ml</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 107px;">Bba-K1614019</td>
 +
<td style="width: 44px;">Can</td>
 +
<td style="width: 86px;">Centrifuged</td>
 +
<td style="width: 73px;">&gt;300</td>
 +
<td style="width: 101px;">&nbsp; &nbsp;-</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 107px;">Bba-K1499004&nbsp;&nbsp;&nbsp;&nbsp;</td>
 +
<td style="width: 44px;">Can</td>
 +
<td style="width: 86px;">0x</td>
 +
<td style="width: 73px;">33</td>
 +
<td style="width: 101px;">3*10^2 /ml</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 107px;">Bba-K1499004&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</td>
 +
<td style="width: 44px;">Can</td>
 +
<td style="width: 86px;">Centrifuged</td>
 +
<td style="width: 73px;">228</td>
 +
<td style="width: 101px;">&nbsp; &nbsp;-&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 107px;">Positive controle: Bba- K1499004</td>
 +
<td style="width: 44px;">None</td>
 +
<td style="width: 86px;">0x</td>
 +
<td style="width: 73px;">&gt; 300</td>
 +
<td style="width: 101px;">&nbsp; &nbsp;-</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 107px;">Negative controle: outgrow medium</td>
 +
<td style="width: 44px;">none</td>
 +
<td style="width: 86px;">0x</td>
 +
<td style="width: 73px;">18</td>
 +
<td style="width: 101px;">&nbsp; &nbsp;-&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p>Tabel 1: results from the transformation with the biobricks Bba-K1614019 and Bba-K1499004.&nbsp;<br>
 +
<br>
 +
The plates show CFU which can be used for further experiments. &nbsp;The negative controle also has CFU, this shows that the outgrow medium was infected and not sterile.&nbsp;<br>
 +
The plates are stored by 4&deg;C.&nbsp;<br>
 +
<br>
 +
Conclusion:&nbsp;<br>
 +
<br>
 +
The transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4&deg;C and can be used for further experiments.&nbsp;<br>
 +
The CFU from the&nbsp;negative controle will be plated on a plate with antibiotics to make sure there isn't a plasmid in the bacteria. The outgrow medium will be sterilized by filter&nbsp;again.&nbsp;<br>
 +
<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing different ATP sensors</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
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 +
<h2 class="date" style="color: white;">July 24</h2>
 +
</div>
 +
 +
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 +
#switch-58:checked ~ #card-58 {
 +
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 +
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 +
}
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#switch-58:checked ~ #card-58 .entryform {
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#switch-58:checked ~ #card-58 .cardform,
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 +
#switch-58:checked ~ #card-58::after {
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display: none;
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</style>
 +
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<input type="checkbox" id="switch-58" class="switch"></input>
 +
<div class="card software-card" id="card-58" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Notebook generator</h1>
 +
<h3 class="date">July 25</h3>
 +
<label class="back" for="switch-58">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Automatic generation of entries completed
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook generator</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-58" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 25</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-59:checked ~ #card-59 {
 +
width: 100%;
 +
height: auto;
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}
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#switch-59:checked ~ #card-59 .entryform {
 +
display: block;
 +
}
 +
#switch-59:checked ~ #card-59 .cardform,
 +
#switch-59:checked ~ #card-59::before,
 +
#switch-59:checked ~ #card-59::after {
 +
display: none;
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}
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</style>
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<input type="checkbox" id="switch-59" class="switch"></input>
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<div class="card software-card" id="card-59" onclick="">
 +
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<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Google Drive API</h1>
 +
<h3 class="date">July 25</h3>
 +
<label class="back" for="switch-59">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Used the Google drive REST API to download the entries from drive
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Google Drive API</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-59" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 25</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-60:checked ~ #card-60 {
 +
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 +
height: auto;
 +
}
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#switch-60:checked ~ #card-60 .entryform {
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 +
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 +
#switch-60:checked ~ #card-60 .cardform,
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#switch-60:checked ~ #card-60::before,
 +
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display: none;
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 +
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<input type="checkbox" id="switch-60" class="switch"></input>
 +
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 +
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 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Automatic uploader</h1>
 +
<h3 class="date">July 25</h3>
 +
<label class="back" for="switch-60">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Used the igemwiki API to update pages using software
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Automatic uploader</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-60" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 25</h2>
 +
</div>
 +
 +
</div><style>
 +
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 +
width: 100%;
 +
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 +
}
 +
#switch-61:checked ~ #card-61 .entryform {
 +
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 +
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 +
#switch-61:checked ~ #card-61 .cardform,
 +
#switch-61:checked ~ #card-61::before,
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 +
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 +
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 +
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 +
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<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">July 27</h3>
 +
<label class="back" for="switch-61">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>For further experiments there is&nbsp;isolated DNA needed of the biobricks J23100, K133071, K173003 and I134353. The DNA is isolated out of the bacteria with the help of a plasmid purifaction kit. After isolation this DNA can be used for digestion and ligation or other experiments.&nbsp;<span style="font-size:12px;">J23100 (from the glycerolstock): 325,59 ng/ul, K173003: 217,06 ng/ul, K133071: 186,79 ng/ul,&nbsp;</span>I13453: 88,18 ng/ul</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:12px;"><strong>Experiment 3: Minipreps (DNA isolation)</strong></span><br>
 +
27-07-18<br>
 +
<br>
 +
<span style="font-size:12px;">Minipreps of the following biobricks in NEB10B&egrave;ta:</span><br>
 +
-J23100: Constitutive promoter<br>
 +
-K133071: Urea --&gt; ammonia + CO2<br>
 +
-K173003: Pyruvate --&gt; acetaldehyde + CO2<br>
 +
-I134353: Promoter (AraC protein binds with arabinose)<br>
 +
<br>
 +
<span style="font-size:12px;"><strong>Materials&nbsp;</strong></span><br>
 +
- Mini prep, Plasmid Purification Kit, Machery Nagel&nbsp;<br>
 +
- Eppendorf cups<br>
 +
- Variable Volume Pipets<br>
 +
- sterile pipet points<br>
 +
-Chloramphenicol (40 mg/mL)<br>
 +
<br>
 +
<span style="font-size:12px;"><strong>Methods</strong></span><span style="font-size:14px;"><strong>&nbsp;</strong></span><br>
 +
<span style="font-size:12px;"><strong>Miniprep protocol&nbsp;Plasmid Purification Kit, Machery Nagel</strong></span><br>
 +
After the transformation we made an overnight culture from the colonies. The colonies were anted into 20 ml LB and incubated overnight at&nbsp;37&deg;C and 150 rpm. The culture was mini prepped the next day. For isolation about 6 mL overnight culture was used.</p>
 +
 +
<p><meta charset="utf-8"></p>
 +
 +
<p dir="ltr"><br>
 +
<b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">Plasmid DNA purification</b>&nbsp;(<b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">NucleoSpin� Plasmid EasyPure)<br>
 +
Machery Nagel Mini preps</b><br>
 +
&nbsp;</p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">1 Cultivate</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">and harvest bacterial cells</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">12,000 x g, 30 s</b><br>
 +
&nbsp;</p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">2 Cell lysis 150 &mu;L Buffer A1</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">250 &mu;L Buffer A2</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">RT, up to 2 min</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">350 &mu;L Buffer A3</b></p>
 +
 +
<p dir="ltr"><br>
 +
<b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">3 Clarification of the lysate</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">&gt; 12,000 x g, 3 min</b></p>
 +
 +
<p dir="ltr"><br>
 +
<b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">4 Bind DNA</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">Load supernatant</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">1,000�2,000 x g, 30 s</b></p>
 +
 +
<p dir="ltr"><br>
 +
<b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">5 Wash and dry silica</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">membrane</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">450 &mu;L Buffer AQ</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">&gt; 12,000 x g, 1 min</b></p>
 +
 +
<p dir="ltr"><br>
 +
<b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">6 Elute DNA 50 &mu;L Buffer AE</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">RT, 1 min</b></p>
 +
 +
<p dir="ltr"><b id="docs-internal-guid-a3024d5d-7fff-fe59-d501-6ae2947d0ae1">&gt; 12,000 x g, 1 min</b></p>
 +
 +
<p><br>
 +
<span style="font-size:12px;"><strong>Results</strong></span><br>
 +
<span style="font-size:12px;">J23100: 325,59 ng/ul<br>
 +
K173003: 217,06 ng/ul<br>
 +
K133071: 186,79 ng/ul</span><br>
 +
I13453: 88,18 ng/ul</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/a/a5/T--Rotterdam_HR--nanodrop27_07.jpeg');"></div>
 +
<p>Figure 1: Minipreps 27-07-18</p>
 +
  </div>
 +
  </div>  
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
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</svg>
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<label for="switch-61" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">July 27</h2>
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</svg>
 +
<h1 class="title">Assessing different ATP sensors</h1>
 +
<h3 class="date">July 27</h3>
 +
<label class="back" for="switch-62">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>To be able to always get the necessary biobricks, we made an glycerol stock after every transformation. The glycerol stocks are stored at -80 degrees Celsius. If needed, they can retrieved from this storage to use for experiments.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p>Making glycerol stocks of the biobricks.&nbsp;<br>
 +
<br>
 +
Materials:&nbsp;<br>
 +
- 87% glycerol&nbsp;<br>
 +
- culture with biobrick plasmids<br>
 +
- LB-medium<br>
 +
- Variable Volume Pipette<br>
 +
- Sterile pipetpoints<br>
 +
<br>
 +
<br>
 +
Method:&nbsp;<br>
 +
<br>
 +
The colonies from the transformation were used to make an overnight culture. The culture was made by putting a colony in LB-medium and then the culture was&nbsp;incubated for a night at 37&deg;C. &nbsp;<br>
 +
<br>
 +
The next day the cultures had grown close and were ready for a glycerol stock. For making glycerol stocks there was added 250 ul 87% glycerol to 1 ml culture. This was frozen in -80&deg;C. &nbsp;<br>
 +
<br>
 +
The stocks were stored at -80&deg;C in drawer 3, tower 1, drawer&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing different ATP sensors</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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</svg>
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<h1 class="title">Automation program</h1>
 +
<h3 class="date">July 29</h3>
 +
<label class="back" for="switch-63">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Connected the page uploader to the notebook generator
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Automation program</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<h1 class="title">Automatic uploader</h1>
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<h3 class="date">July 29</h3>
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<label class="back" for="switch-64">&times;</label>
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</div>
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<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Created a program that uploads all updated pages
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Automatic uploader</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">July 29</h3>
 +
<label class="back" for="switch-65">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
After considering multiple tactics, we've decided to follow the standard iGEM 3A assembly in our following cloning steps. To this end we started with an overnight digest of all the different parts.
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 800px;">
 +
<tbody>
 +
<tr>
 +
<td>BioBrick BBa_ code</td>
 +
<td>Restriction enzyme 1</td>
 +
<td>Restriction enzyme 2</td>
 +
</tr>
 +
<tr>
 +
<td>J23100</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>J23105</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>J23113</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>B0030</td>
 +
<td>EcoRI</td>
 +
<td>XbaI</td>
 +
</tr>
 +
<tr>
 +
<td>B0031</td>
 +
<td>EcoRI</td>
 +
<td>XbaI</td>
 +
</tr>
 +
<tr>
 +
<td>B0032</td>
 +
<td>EcoRI</td>
 +
<td>XbaI</td>
 +
</tr>
 +
<tr>
 +
<td>K352001</td>
 +
<td>SpeI</td>
 +
<td>PstI</td>
 +
</tr>
 +
<tr>
 +
<td>B0015</td>
 +
<td>XbaI</td>
 +
<td>PstI</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p class="experiment"></p>
 +
<p>Used&nbsp;pipette scheme from protocol 7 to prepare the DNA to be digested with multiple combinations of EcoRI-HF, SpeI-HF, PstI and XbaI.</p>
 +
With this we started a digest of the following BioBricks using the following Restriction enzyme couples:
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 800px;">
 +
<tbody>
 +
<tr>
 +
<td>BioBrick BBa_ code</td>
 +
<td>Restriction enzyme 1</td>
 +
<td>Restriction enzyme 2</td>
 +
</tr>
 +
<tr>
 +
<td>J23100</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>J23105</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>J23113</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>B0030</td>
 +
<td>EcoRI</td>
 +
<td>XbaI</td>
 +
</tr>
 +
<tr>
 +
<td>B0031</td>
 +
<td>EcoRI</td>
 +
<td>XbaI</td>
 +
</tr>
 +
<tr>
 +
<td>B0032</td>
 +
<td>EcoRI</td>
 +
<td>XbaI</td>
 +
</tr>
 +
<tr>
 +
<td>K352001</td>
 +
<td>SpeI</td>
 +
<td>PstI</td>
 +
</tr>
 +
<tr>
 +
<td>B0015</td>
 +
<td>XbaI</td>
 +
<td>PstI</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<br>
 +
Digestion was started on the 30th of July, ~16:00, 37&deg;C and 185 RPM.<br>
 +
Digestion was stopped on the 31st of July, ~10:00.
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>09</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-65" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 29</h2>
 +
</div>
 +
 +
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 +
#switch-66:checked ~ #card-66 {
 +
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 +
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 +
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<input type="checkbox" id="switch-66" class="switch"></input>
 +
<div class="card wetlab-card" id="card-66" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Making competent NEB10beta cells</h1>
 +
<h3 class="date">July 30</h3>
 +
<label class="back" for="switch-66">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Jos Veldscholte</div>
 +
<div class="description">
 +
<p>Made stock solution of MgSO<sub>4</sub> and pure culture of NEB10 beta cells</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Making 1 M sterile MgSO<sub>4</sub></strong><br>
 +
&asymp; 14.00: Dissolved 12,32 g MgSO<sub>4</sub> in 50 mL demi water en filtered through 0.22&nbsp;&mu;m filter.<br>
 +
<br>
 +
<strong>Making pure culture of NEB10 beta cells</strong><br>
 +
&asymp;15.00:<strong>&nbsp;</strong>20 mL LB agar + 10&nbsp;&mu;L tetracyclin (50 mg/mL) to pour 1 plate.<br>
 +
16:30: streaked bacteria from glycerol stock: -80, section III, tower 1, shelf IV, box III, 6B<br>
 +
Left in 37&deg;C room for o.n. growth.</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Making competent NEB10beta cells</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-66" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 30</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-67:checked ~ #card-67 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-67:checked ~ #card-67 .entryform {
 +
display: block;
 +
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 +
#switch-67:checked ~ #card-67 .cardform,
 +
#switch-67:checked ~ #card-67::before,
 +
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 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-67" class="switch"></input>
 +
<div class="card software-card" id="card-67" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Page generator</h1>
 +
<h3 class="date">July 31</h3>
 +
<label class="back" for="switch-67">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Built a page builder that automatically adds the header and footer before uploading
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Page generator</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-67" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 31</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-68:checked ~ #card-68 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-68:checked ~ #card-68 .entryform {
 +
display: block;
 +
}
 +
#switch-68:checked ~ #card-68 .cardform,
 +
#switch-68:checked ~ #card-68::before,
 +
#switch-68:checked ~ #card-68::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-68" class="switch"></input>
 +
<div class="card software-card" id="card-68" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Google Drive API</h1>
 +
<h3 class="date">July 31</h3>
 +
<label class="back" for="switch-68">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Redone the google API handler for reusability and readability and made the google API handler generic
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Google Drive API</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-68" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 31</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-69:checked ~ #card-69 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-69:checked ~ #card-69 .entryform {
 +
display: block;
 +
}
 +
#switch-69:checked ~ #card-69 .cardform,
 +
#switch-69:checked ~ #card-69::before,
 +
#switch-69:checked ~ #card-69::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-69" class="switch"></input>
 +
<div class="card software-card" id="card-69" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Automation program</h1>
 +
<h3 class="date">July 31</h3>
 +
<label class="back" for="switch-69">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
revamped the main program
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Automation program</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-69" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 31</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-70:checked ~ #card-70 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-70:checked ~ #card-70 .entryform {
 +
display: block;
 +
}
 +
#switch-70:checked ~ #card-70 .cardform,
 +
#switch-70:checked ~ #card-70::before,
 +
#switch-70:checked ~ #card-70::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-70" class="switch"></input>
 +
<div class="card wetlab-card" id="card-70" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">July 31</h3>
 +
<label class="back" for="switch-70">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>Though we had started with our ligation using the rAPId dephos and ligation kit, we'd simultaniously put our DNA on gel, later revealing the digest to not have been succesful. Carefull examination of our work made us realise the used restriction enzyme dilutions to not have been made correctly.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Using the rAPid Dephos and Ligation Kit we performed a dephosphorilation and ligation of our DNA.<br>
 +
After checking our gel with our digestion products we later&nbsp;determined that the digestion didn't go as expected<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>10</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-70" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 31</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-71:checked ~ #card-71 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-71:checked ~ #card-71 .entryform {
 +
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#switch-71:checked ~ #card-71 .cardform,
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#switch-71:checked ~ #card-71::before,
 +
#switch-71:checked ~ #card-71::after {
 +
display: none;
 +
}
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</style>
 +
 +
<input type="checkbox" id="switch-71" class="switch"></input>
 +
<div class="card wetlab-card" id="card-71" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Making competent NEB10beta cells</h1>
 +
<h3 class="date">July 31</h3>
 +
<label class="back" for="switch-71">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Jos Veldscholte</div>
 +
<div class="description">
 +
<p>Made Ksi broth (for competent cells) and started o.n. cultures</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Make Ksi broth (for competent cells)</strong><br>
 +
Dissolved 20 g Tryptone (Lab M limited) and 5 g Yeast extract&nbsp;(Lab M limited) in 1 liter demi water, added 10 ml 1 M NaCl and adjusted pH to 7.5 with KOH.<br>
 +
Autoclaved.<br>
 +
16:30<br>
 +
2 tubes with 2.5 mL Ksi inoculated with 1 colony NEB10beta from yesterday's plate. (Added 50 microliters of 1 M MgSO4 at 16:30 becauce I forgot to ad it to de autoclaved medium.&nbsp;<br>
 +
Added 10 mL of 1 M MgSO4 to each half liter autoclaved medium (=Ksi).</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Making competent NEB10beta cells</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-71" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">July 31</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-72:checked ~ #card-72 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-72:checked ~ #card-72 .entryform {
 +
display: block;
 +
}
 +
#switch-72:checked ~ #card-72 .cardform,
 +
#switch-72:checked ~ #card-72::before,
 +
#switch-72:checked ~ #card-72::after {
 +
display: none;
 +
}
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</style>
 +
 +
<input type="checkbox" id="switch-72" class="switch"></input>
 +
<div class="card software-card" id="card-72" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<h1 class="title">Automation program</h1>
 +
<h3 class="date">August 1</h3>
 +
<label class="back" for="switch-72">&times;</label>
 +
</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Finished redoing the application for generating and uploading pages
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Automation program</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
 +
</svg>
 +
<label for="switch-72" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">August 1</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-73:checked ~ #card-73 {
 +
width: 100%;
 +
height: auto;
 +
}
 +
#switch-73:checked ~ #card-73 .entryform {
 +
display: block;
 +
}
 +
#switch-73:checked ~ #card-73 .cardform,
 +
#switch-73:checked ~ #card-73::before,
 +
#switch-73:checked ~ #card-73::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-73" class="switch"></input>
 +
<div class="card wetlab-card" id="card-73" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Making competent NEB10beta cells</h1>
 +
<h3 class="date">August 1</h3>
 +
<label class="back" for="switch-73">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Jos Veldscholte</div>
 +
<div class="description">
 +
<p>Made large NEB10 beta culture and aliquoted in 50 microliter portions and snap froze in dry ice/isopropanol</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Making competent NEB10 beta cells</strong><br>
 +
8:45:<br>
 +
Both cultures were fine.<br>
 +
Transfered one culture completely to 100 ml Erlenmeyer with 25 mL Ksi (RT). In shaking incubator at 37&deg; C at 200 rpm.<br>
 +
Filled three one liter Erlenmeyers with 175 mL Ksi and put in shaking incubator in 37&deg; C room to adjust temperature.<br>
 +
25 ml culture: at 8:40 OD<sub>600</sub> was&nbsp;0.64.<br>
 +
Transferred 8 ml to each 175 mL Erlenmeyer. Incubate at&nbsp; 37&deg; C shaking rigorously (no speed indicator).<br>
 +
9:40: OD<sub>600</sub> = 0.-9<br>
 +
10:30: OD<sub>600</sub> = 0.30<br>
 +
10:50 OD<sub>600</sub> = 0.46<br>
 +
10:58 OD<sub>600</sub> = 0.55<br>
 +
Tranferred all cultures to 10 50 mL tubes and tare in pairs.<br>
 +
On ice 11:15<br>
 +
11:35: in Sorvall 4500 rpm, 15&nbsp;min 0&deg; C.<br>
 +
After the run, fluid was on the rotor and in the rotor holes.<br>
 +
Cleaned with lab desinfectant. Rinsed with water and dried with tissue.<br>
 +
From here on everything in the cold room with tubes on ice:<br>
 +
Resuspended pellets in 16,6&nbsp;ml ice&nbsp;cold TFB1 (protocol "competente-cellen-maken"l) each with 10 mL plastic pipet and pipetboy.&nbsp;<br>
 +
Combined into four&nbsp;50 mL tubes. Centrifuged&nbsp;15 min at 4500 rpm.<br>
 +
Resuspended pellets in 60 mL TFB1, kept on ice for 20 min and centrifuged 15 min 4500 rpm.<br>
 +
Respuspended pellets in 10,5 ml TFB2&nbsp;(protocol "competente-cellen-maken"l). Left on Ice for 10 min.<br>
 +
Aliquotted into 50&nbsp;&mu;L portions in 2 mL conical tubes and snap froze in isopropanol on dry ice.<br>
 +
Transferred to -80&deg;C Freezer.</p>
 +
 +
</div>
 +
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<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Making competent NEB10beta cells</h3>
 +
<svg viewBox="0 0 24 24">
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<h2 class="date" style="color: white;">August 1</h2>
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<h1 class="title">Notebook</h1>
 +
<h3 class="date">August 2</h3>
 +
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</div>
 +
<div class="attendees">Rens Boeser</div>
 +
<div class="description">
 +
Added month categorization to the page
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
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<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Notebook</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M14.6,16.6L19.2,12L14.6,7.4L16,6L22,12L16,18L14.6,16.6M9.4,16.6L4.8,12L9.4,7.4L8,6L2,12L8,18L9.4,16.6Z"></path>
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<h2 class="date" style="color: white;">August 2</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">August 2</h3>
 +
<label class="back" for="switch-75">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>We've redone the work mentioned in the previous CooA entry, though today our gel electrophoresis revealed we were only partially succesful, as only the digests with EcoRI and SpeI seem to&nbsp;be&nbsp;correct.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Experiment 9 and 10 have&nbsp;been repeated with a new enzyme dilution.<br>
 +
7th of august: review of the digestions on gel reveal that only the digestions with EcoRI-HF and SpeI-HF worked, while the PstI and XbaI digestion did not retrieve the expected results</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>11</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 2</h2>
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<h1 class="title">Making competent NEB10beta cells</h1>
 +
<h3 class="date">August 2</h3>
 +
<label class="back" for="switch-76">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Jos Veldscholte</div>
 +
<div class="description">
 +
<p>Tested competence by transformation of pUC19</p>
 +
 +
<p class="experiment"></p>
 +
<p>Test sample from NEB&nbsp;competent cells contains 50 pg pUC19 per&nbsp;&mu;L.<br>
 +
Used 2&nbsp;&mu;L per transformation (= 100 ng).<br>
 +
Took three tubes from one of the two boxes.<br>
 +
A 100 ng pUC19<br>
 +
B 100 ng pUC19<br>
 +
C nothing<br>
 +
30 min on ice (11:00)<br>
 +
A 30 s heatshock in waterbath (11:30)<br>
 +
B 90 s heatshock in waterbath (11:30)<br>
 +
C was kept on ice still.<br>
 +
5 min on ice.<br>
 +
Added 1 ml of LB (11:40)<br>
 +
60 min shaking in 37&nbsp;&deg;C room (shaking slowly)<br>
 +
<br>
 +
Poured LB agar plates with 50&mu;g/ml Ampicillin and one plate without antibiotic.<br>
 +
<br>
 +
12:40<br>
 +
Added 1&nbsp;&mu;L of culture A or B to 99&nbsp;&mu;L LB (A4 and B4)<br>
 +
Added 10&nbsp;&mu;L of culture A or B to 90&nbsp;&mu;L LB (A3 and C3)<br>
 +
Plated A3 and 4 and C3 and 4 on LB agar amp.<br>
 +
Plated 100&nbsp;&mu;L A, B and C (A2, B2 and C2) on LB agar amp.<br>
 +
Plated 100&nbsp;&mu;L C on plate without antibiotics (to test whether cells are alive)<br>
 +
Centrifuged&nbsp;A and B 6000 rpm in microcentrifuge for 2 min.<br>
 +
Aspirated supernatant.<br>
 +
Vortexed bacteria in remainder of the medium (&gt;100&nbsp;&mu;L)<br>
 +
Plated on LB agar amp. plates (A1 and B1).<br>
 +
<br>
 +
Consequently:<br>
 +
A was heat shocked for 30 s.<br>
 +
B was heat shocked for 90 s.<br>
 +
1 = 90% of the transformation, 2 = 10%, 3 = 1% and 4 = 0,1%<br>
 +
<br>
 +
Put in automated incubator. Not sure about whether the sytem wil work. Is it on?<br>
 +
Colonies to be counted next monday by Dustin.<br>
 +
If there are no colonies visible, please extend incubation time!</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Making competent NEB10beta cells</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 2</h2>
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 +
<h1 class="title">Making competent NEB10beta cells</h1>
 +
<h3 class="date">August 3</h3>
 +
<label class="back" for="switch-77">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Jos Veldscholte</div>
 +
<div class="description">
 +
<p>Counted colonies. Result: no transformation.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Colonies where counted at ~10:00. Only plate A1 showed signs of colonies (~15). All the others where empty.<br>
 +
After leaving the cells in the incubator for an extra half a day, there where no new colonies.</p>
 +
 +
</div>
 +
</div>
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<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Making competent NEB10beta cells</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 3</h2>
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</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">August 4</h3>
 +
<label class="back" for="switch-78">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
The previous two CooA entries have been repeated, though this time we remade the PstI and XbaI enzyme dilutions. This time the digests where as expected.
 +
<p class="experiment"></p>
 +
<p>Experiment 11&nbsp;has been repeated, but with new dilutions of PstI and XbaI.<br>
 +
After the incubation it was noted that the cup containging BBa_B0015 was almost empty, so it will have to be digested again. (did not happen) --&gt; in the end we had enough<br>
 +
<br>
 +
13th of august --&gt; dephos and ligation was peformed on the obtained biobrick combinations following protocol 8<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
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<div class="cardform">
 +
<div class="number"><b>12</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Making competent NEB10beta cells</h1>
 +
<h3 class="date">August 7</h3>
 +
<label class="back" for="switch-79">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Jos Veldscholte</div>
 +
<div class="description">
 +
<p>Repeated transformation.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Part 4 has been repeated, repeating the same steps but adding two extra positive controlls.</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>06</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Making competent NEB10beta cells</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<label for="switch-79" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">August 7</h2>
 +
</div>
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</div><style>
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</svg>
 +
<h1 class="title">Assessing different ATP sensors</h1>
 +
<h3 class="date">August 9</h3>
 +
<label class="back" for="switch-80">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>For further experiments we needed isolated DNA. The DNA is isolated out of the bacteria with the help of a plasmid purifaction kit. After isolation this DNA can be used for digestion and ligation or other experiments.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Materials:&nbsp;<br>
 +
- Mini prep, Plasmid Purification Kit, Machery Michels&nbsp;<br>
 +
- Eppendorf cups<br>
 +
- Variable Volume Pipets<br>
 +
- sterile pipet points<br>
 +
<br>
 +
<br>
 +
Method:&nbsp;<br>
 +
After the transformation we made an overnight culture from the colonies. The colonies were anted into 20 ml LB and incubated overnight at&nbsp;<span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">37&deg;C and 150 rpm. The culture was mini prepped the next day. For isolation, 5,4 ml was added step wise in a 2ml Eppendorf cup. The culture was then centrifuged at &gt;12000x g for 30 seconds. The supernatant was deposed, and the pellet was resupended with A1 buffer and vortexed. To the resuspended culture was then.&nbsp;</span></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing different ATP sensors</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<label for="switch-80" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">August 9</h2>
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</div>
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<div class="card wetlab-card" id="card-81" onclick="">
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 10</h3>
 +
<label class="back" for="switch-81">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>The biobricks&nbsp;J23100, K133071, K173003 and I134353 were&nbsp;sucessfully digested after the second time. After the&nbsp;digestions the&nbsp;biobricks K133071 and K173003&nbsp;were dephosphorylated and ligated with the inserts J23100 and I13453. This was done in the original backbone of K133071 and K173003, and not another control backbone. To know&nbsp;if the biobricks were right ligated this was done by testing practically.&nbsp;See the experiments:&nbsp;<em>Testing gas production.</em>&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 14px;"><strong>Experiment 4: Digestion, Defosforylation and&nbsp;ligation</strong></span><br>
 +
<span style="font-size:14px;"><strong>Digestions</strong></span><br>
 +
<span style="font-size:12px;">30-07-18 and 08-08-18<br>
 +
<br>
 +
Biobricks: J23100, K133071, K173003 and I13453</span><br>
 +
<br>
 +
Used&nbsp;pipette scheme (Table 1) to prepare the DNA to be cut with multiple combinations of EcoRI-HF, SpeI-HF, PstI and XbaI.</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td>DNA</td>
 +
<td>~ 2 &mu;g</td>
 +
</tr>
 +
<tr>
 +
<td>NEBbuffer 2.1</td>
 +
<td>5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>Fill to 100 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>(Table 1 ; Double Digest Pipette Scheme using :&nbsp;<a href="https://nebcloner.neb.com/#!/redigest"><u><font color="#0066cc">https://nebcloner.neb.com/#!/redigest</font></u></a>, and doubling the amount)</em><br>
 +
<br>
 +
<br>
 +
<strong>Biobricks:</strong><br>
 +
J23100: Constitutive promotor<br>
 +
K133071: Urea --&gt; ammonia + CO2<br>
 +
K173003: Pyruvate --&gt; acetaldehyde + CO2<br>
 +
I134353: Promotor (AraC protein binds with arabinose)<br>
 +
<br>
 +
With this we started a digest of the following BioBricks using the following Restriction enzyme couples (Table 2)<br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 571px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 87px;"><strong>Number</strong></td>
 +
<td style="width: 162px;"><strong>BioBrick BBa_ code</strong></td>
 +
<td style="width: 191px;"><strong>Restriction enzyme 1</strong></td>
 +
<td style="width: 198px;"><strong>Restriction enzyme 2</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 87px;">9</td>
 +
<td style="width: 162px;">J23100 (35 bp)</td>
 +
<td style="width: 191px;">EcoRI</td>
 +
<td style="width: 198px;">SpeI</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 87px;">11</td>
 +
<td style="width: 162px;">K133071 (1707 bp)</td>
 +
<td style="width: 191px;">EcoRI</td>
 +
<td style="width: 198px;">XbaI</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 87px;">12</td>
 +
<td style="width: 162px;">K173003 (3052 bp)</td>
 +
<td style="width: 191px;">EcoRI</td>
 +
<td style="width: 198px;">XbaI</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 87px;">10</td>
 +
<td style="width: 162px;">I13453 (130 bp)</td>
 +
<td style="width: 191px;">EcoRI</td>
 +
<td style="width: 198px;">SpeI</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><em>Table 2: Biobricks and restriction enzymes</em><br>
 +
<br>
 +
Digestion was started on the 30th of July, ~16:00, 37&deg;C and 185 RPM.<br>
 +
Digestion was stopped on the 31st of July, ~10:00.<br>
 +
<br>
 +
This has been done in tandem with the corresponding part from&nbsp;<a href="/members/experiments/browser/#view=experiment&amp;nodeID=313406"><u><font color="#0066cc">Cloning Mulitple BioBricks to Assess CooA Production</font></u></a>&nbsp;Part X&nbsp;</p>
 +
 +
<p><br>
 +
<br>
 +
<strong><span style="font-size:14px;">Dephosphorylation</span></strong><br>
 +
<span style="font-size:12px;">10-08-18</span><br>
 +
<br>
 +
K133071 and K173003 were dephosphoralized:<br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 313px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 128px;"><strong>Volume</strong></td>
 +
<td style="width: 172px;"><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">25&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;">Vector DNA (<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">0,5&nbsp;&mu;g)</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">3&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">phosphatase buffer (10x)</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">1&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">milli Q</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">1&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">phosphatase (1 U/&mu;L)</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">30&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Total</span></span></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<div class="step" data-stepid="362681">
 +
<div class="stepBody"><em>Table 3: Pipetting scheme</em><br>
 +
<br>
 +
The new DNA concentration of the reaction mix is 0,5&nbsp;&mu;g/30&nbsp;&mu;L<br>
 +
The reaction mix was incubated for 10 minutes at 37&nbsp;&deg;C<br>
 +
The phosphatase was inactivated for 2 minutes at 75&nbsp;&deg;C</div>
 
</div>
 
</div>
  
−
<!-- ------ CONTENT END  ------ -->
+
<div class="step" data-stepid="362684">
 +
<div class="stepHeader">
 +
<div class="stepTitle">&nbsp;</div>
  
 +
<div class="stepMenu">
 +
<ul>
 +
<li class="btnEditSave" style="display: none;">&nbsp;</li>
 +
<li class="btnMoveStep" style="display: none;">&nbsp;</li>
 +
<li class="btnRemoveStep warning" style="display: none;">&nbsp;</li>
 +
</ul>
 +
</div>
 +
</div>
 +
 +
<div class="stepBody">
 +
<div class="stepContents">For the ligation mix 100 ng of vector DNA was used from the dephosphorylation reaction mix.<br>
 +
The following vector-insert combinations were ligated:</div>
 +
</div>
 +
</div>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 403px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 119px;"><strong>Vector</strong></td>
 +
<td style="width: 132px;"><strong>Insert 1</strong></td>
 +
<td style="width: 134px;"><strong>Insert 2</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 119px;">K133071</td>
 +
<td style="width: 132px;">J23100</td>
 +
<td style="width: 134px;">I13453</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 119px;">K173003</td>
 +
<td style="width: 132px;">J23100</td>
 +
<td style="width: 134px;">I13453</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<div class="stepMenu">
 +
<ul>
 +
<li class="btnEditSave" style="display: none;">&nbsp;</li>
 +
<li class="btnMoveStep" style="display: none;">&nbsp;</li>
 +
<li class="btnRemoveStep warning" style="display: none;">&nbsp;</li>
 +
</ul>
 +
</div>
 +
<em>Table 4: Biobrick combination</em><br>
 +
<br>
 +
The original plasmids of the vector and inserts that were digested are about the same size in bp and the ratio vector DNA-insert DNA is determined to be 1:3. Therefore, the amount of insert DNA that was used for the ligation was&nbsp;140 ng (rounded up to 7&nbsp;&mu;L of the digestion reaction mix). Because there was only 5&nbsp;&mu;L left of the K352001 insert DNA, the total 5&nbsp;&mu;L were used and an extra 2&nbsp;&mu;L of milli Q water were added to the pre-ligation mix.<br>
 +
<br>
 +
Pre-Ligationmix:
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 313px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 128px;"><strong>Volume</strong></td>
 +
<td style="width: 172px;"><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">6 &mu;L</span></span></td>
 +
<td style="width: 172px;">Vector DNA (<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">0,5&nbsp;&mu;g/30 &mu;L)</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">7 &mu;L</span></span></td>
 +
<td style="width: 172px;"><font face="arial, helvetica, sans-serif">Insert DNA (2&nbsp;</font><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">&mu;g/100&nbsp;&mu;L)</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">4 &mu;L</span></span></td>
 +
<td style="width: 172px;">Delutionbuffer (5x)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">3 &mu;L</span></span></td>
 +
<td style="width: 172px;">Milli Q</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">20&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Total</span></span></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 5: Pre-ligationmix pipette scheme</em><br>
 +
<br>
 +
Ligationmix:<br>
 +
incubation at room-temperature for 20 minutes.
 +
<div class="stepBody">
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 313px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 128px;"><strong>Volume</strong></td>
 +
<td style="width: 172px;"><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">10 &mu;L</span></span></td>
 +
<td style="width: 172px;">Pre-ligationmix</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">2 &mu;L</span></span></td>
 +
<td style="width: 172px;">10x T4 ligationbuffer</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">1 &mu;L</span></span></td>
 +
<td style="width: 172px;">T4 DNA ligase (1U/<span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">&mu;L)</span></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">8 &mu;L</span></span></td>
 +
<td style="width: 172px;">Milli Q</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 128px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">20&nbsp;&mu;L</span></span></td>
 +
<td style="width: 172px;"><span style="font-size:12px;"><span style="font-family:arial,helvetica,sans-serif;">Total</span></span></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 6: Ligationmix pipette scheme</em><br>
 +
&nbsp;</div>
 +
 +
<div class="stepBody">
 +
<div class="step" data-stepid="362684">
 +
<div class="stepBody">
 +
<div class="stepContents">All the vectors contain the CamR gene. 30 plates were poured with Cam (14 for the transformed cells (100&mu;L), 14 for the transformed cells (900&nbsp;&mu;L centrifuged and resuspended in 100&nbsp;&mu;L and 1 positive and 1 negative control)</div>
 +
</div>
 +
</div>
 +
</div>
 +
 +
<p>&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 10</h2>
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 +
</svg>
 +
<h1 class="title">Urea and sodium pyruvate test for resistance E.coli</h1>
 +
<h3 class="date">August 10</h3>
 +
<label class="back" for="switch-82">&times;</label>
 +
</div>
 +
<div class="attendees">Randall de Waard | Elise Grootscholten</div>
 +
<div class="description">
 +
<p><span style="font-size:14px;">Testing different amounts of urea and sodiumpyruvate to know which concentrations the bacteria survive.</span></p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 16px;"><strong>Experiment 1:&nbsp;Testing different concentrations of Urea and Sodium pyruvate</strong></span><br>
 +
<span style="font-size:14px;"><strong>08-08-18</strong></span><br>
 +
<br>
 +
<span style="font-size:14px;">Testing different amountsof urea and sodiumpyruvate to know which concentrations the bacteria survive.<br>
 +
<br>
 +
-First let the LB agarose heat up (95 degrees Celcius)<br>
 +
-Second, weigh urea and sodium pyruvate in beakers (see table 1)</span></p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 404px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 122px;"><span style="font-size:14px;"><strong>What</strong></span></td>
 +
<td style="width: 113px;"><span style="font-size:14px;"><strong>How much</strong></span></td>
 +
<td style="width: 153px;"><span style="font-size:14px;"><strong>Solution</strong></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 113px;"><span style="font-size:14px;">1 g</span></td>
 +
<td style="width: 153px;"><span style="font-size:14px;">1 mL purified water</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 113px;"><span style="font-size:14px;">2,4 g</span></td>
 +
<td style="width: 153px;"><span style="font-size:14px;">2 mL purified water</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 113px;"><span style="font-size:14px;">4,8 g</span></td>
 +
<td style="width: 153px;"><span style="font-size:14px;">4 mL purified water</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;"><span style="font-size:14px;">Sodium pyruvate </span></td>
 +
<td style="width: 113px;"><span style="font-size:14px;">5,7 mL</span></td>
 +
<td style="width: 153px;"><span style="font-size:14px;">None</span></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><span style="font-size:14px;"><em>Table 1: weighing urea and sodium pyruvate for LB agarose plates.</em><br>
 +
-Purify these substances, so it is free of bacteria. Work&nbsp;beneath&nbsp;the flame and pour it in to plastic 50 mL tubes.<br>
 +
-Pour the LB agarose in the plastic tubes when it's about 45 degrees Celcius and mix gently.<br>
 +
-Pour the LB agarose mix in a petridish and wait until it's dry. Do this for all the plastic tubes.<br>
 +
-Also fill 2 petridishes with LB agarose without other substances (positive and negative control).<br>
 +
-prick a BL21(DE3) bacteria and put it in a 100 mL erlenmeyer with 20 mL culture medium.<br>
 +
<br>
 +
<strong>09-08-18</strong><br>
 +
<br>
 +
Pipette 100 ul overnight culture on each plate,&nbsp;except on&nbsp;the negative control,&nbsp;and divide it.&nbsp;(24h 37 degrees Celcius)</span><br>
 +
<br>
 +
&nbsp;</p>
 +
 +
<p><span style="font-size:14px;"><strong>Results</strong></span><br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 526px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;"><strong>What</strong></span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;"><strong>How much</strong></span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;"><strong>Solution</strong></span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;"><strong>Result</strong></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;">1 g</span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;">1 mL purified water</span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;">Grow</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;">2,4 g</span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;">2 mL purified water</span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;">No grow</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;">4,8 g</span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;">4 mL purified water</span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;">No grow</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;">Sodium pyruvate</span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;">5,7 mL</span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;">Grow</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;">Positive control</span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;">Grow</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 123px;"><span style="font-size:14px;">Negative control</span></td>
 +
<td style="width: 95px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 170px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 117px;"><span style="font-size:14px;">No grow</span></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><span style="font-size:14px;"><em>Table 1: Results experiment 1</em><br>
 +
<br>
 +
<strong>Conclusion</strong><br>
 +
<br>
 +
More tests will be done with urea and sodiumpyruvate because with 1 g urea the bacteria live and with 2,4 g urea the bacteria die.<br>
 +
The bacteria survive 5,7 mL sodiumpyruvate so we will test with higher concentrations.</span><br>
 +
&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Urea and sodium pyruvate test for resistance E.coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 13</h3>
 +
<label class="back" for="switch-83">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Transformations of the biobricks (K133071 + J23100), (K13071 + I1345), (K173003 + J23100) and (K173003 + I13453) in NEB10B&egrave;ta.&nbsp;There was no grow except for the biobrick combination K133071 + J23100.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 16px;"><strong>Experiment 5: Transformation</strong></span></p>
 +
 +
<p><span style="color:#000000;"><span style="font-size:12px;">13-08-2018<br>
 +
Transformations of the ligations of experiment 4</span></span><br>
 +
<br>
 +
<strong style="color: rgb(0, 0, 0); font-family: arial, helvetica, sans-serif;">Materials</strong></p>
 +
 +
<p><span style="font-family:arial,helvetica,sans-serif;"><span style="color:#000000;">-Chloramphenicol (40 mg/mL)<br>
 +
-LB-agar<br>
 +
-Ligations<br>
 +
-Heat block,42 degrees Celcius<br>
 +
-Shaker, 37 degrees Celsius&nbsp;<br>
 +
-Chemo competent cells NEB10b&egrave;ta (New England Biolabs)</span></span><br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
<br>
 +
10 ul ligationmix is used for every transformation</p>
 +
 +
<h1><span style="font-size: 16px;">Transformation Protocol</span><br>
 +
<span style="font-size: 14px;"><span style="color: rgb(0, 0, 0);">Overview</span></span></h1>
 +
 +
<p><span style="font-size: 14px;"><span style="color: rgb(0, 0, 0);"><span style="font-family: arial,helvetica,sans-serif;">Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.</span></span></span></p>
 +
 +
<section class="paragraph  ">
 +
<h2><span style="font-size: 14px;">Protocol</span></h2>
 +
 +
<ol>
 +
<li>Thaw competent cells on ice.</li>
 +
<li>Chill approximately 5 ng (2 &mu;l) of the ligation mixture in a 1.5 ml microcentrifuge tube.</li>
 +
<li>Add 50 &mu;l of competent cells to the DNA. Mix gently by pipetting up and down or flicking the tube 4�5 times to mix the cells and DNA. Do not vortex.</li>
 +
<li>Place the mixture on ice for 30 minutes. Do not mix.</li>
 +
<li>Heat shock at 42&deg;C for 30 seconds*. Do not mix.</li>
 +
<li>Add 950 &mu;l of room temperature media* to the tube.</li>
 +
<li>Place tube at 37&deg;C for 60 minutes. Shake vigorously (250 rpm) or rotate.</li>
 +
<li>Warm selection plates to 37&deg;C.</li>
 +
<li>Spread 50�100 &mu;l of the cells and ligation mixture onto the plates.</li>
 +
<li>Incubate overnight at 37&deg;C.
 +
<p>* Please note: For the duration and temperature of the heat shock step as well as for the media to be used during the recovery period, please follow the recommendations provided by the competent cells� manufacturer.</p>
 +
</li>
 +
</ol>
 +
</section>
 +
 +
<p><br>
 +
Instead of 2 ul of the ligation mix, use 10 ul ligation mix.</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
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 +
<h2 class="date" style="color: white;">August 13</h2>
 +
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 14</h3>
 +
<label class="back" for="switch-84">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>A colony PCR is done for the NEB10B&egrave;ta E.coli cells with expected&nbsp;the biobrick combination of K133071 with J23100. Nevertheless, on a gel the difference with and without promotor couldn't be seen. So there must be another way of proving the right biobricks are there.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 14px;"><strong>Expermiment 6: Colony PCR K133071 + J23100</strong></span><br>
 +
14-08-18<br>
 +
<br>
 +
Mastermix</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 470px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 163px;"><strong>What</strong></td>
 +
<td style="width: 147px;"><strong>1 reaction</strong></td>
 +
<td style="width: 146px;"><strong>30 reactions</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;">Taq buffer(10x)</td>
 +
<td style="width: 147px;">5 ul</td>
 +
<td style="width: 146px;">150 ul</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;">10 mM dntp</td>
 +
<td style="width: 147px;">1 ul</td>
 +
<td style="width: 146px;">30 ul</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;">10 uM Forward primer</td>
 +
<td style="width: 147px;">1 ul</td>
 +
<td style="width: 146px;">30 ul</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;">10 uM Reverse primer</td>
 +
<td style="width: 147px;">1 ul</td>
 +
<td style="width: 146px;">30 ul</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;">Taq polymerase</td>
 +
<td style="width: 147px;">0,25 ul</td>
 +
<td style="width: 146px;">7,5 ul</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;">Nucease free hydrogen</td>
 +
<td style="width: 147px;">41,75 ul</td>
 +
<td style="width: 146px;">1252,5 ul</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;"><strong>Total</strong></td>
 +
<td style="width: 147px;"><strong>50 ul</strong></td>
 +
<td style="width: 146px;"><strong>1500 ul</strong></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p>&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>06</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 14</h2>
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</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 15</h3>
 +
<label class="back" for="switch-85">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
The ligations and transformations of the biobricks K173003 + J23100, K173003 + I13453, and K133071 + I13453 have been performed again.&nbsp;There was a lot of grow on the agar plates. To know&nbsp;if the biobricks were right ligated this was done by testing practically.&nbsp;See the experiments:&nbsp;<em>Testing gas production.</em>&nbsp;
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 14px;"><strong>Experiment 7: Ligation of biobricks</strong></span><br>
 +
15-08-18<br>
 +
<br>
 +
K173003 + J23100<br>
 +
K173003 + I13453<br>
 +
K133071 + I13453<br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 300px;">
 +
<thead>
 +
<tr>
 +
<th scope="col" style="width: 79px;">Volume</th>
 +
<th scope="col" style="width: 213px;">Compound</th>
 +
</tr>
 +
</thead>
 +
<tbody>
 +
<tr>
 +
<td style="width: 67px;">6&nbsp;&mu;L</td>
 +
<td style="width: 201px;">vector DNA (0,5&nbsp;&mu;g/30&nbsp;&mu;L) (100 ng)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">14&nbsp;&mu;L</td>
 +
<td style="width: 201px;">insert DNA (2&nbsp;&mu;g/100 &mu;L) (280 ng)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">4&nbsp;&mu;L</td>
 +
<td style="width: 201px;">dilutionbuffer (5x)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">1&nbsp;&mu;L</td>
 +
<td style="width: 201px;">milli Q water</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">25&nbsp;&mu;L</td>
 +
<td style="width: 201px;">Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><br>
 +
Use 10 mL<br>
 +
<br>
 +
Ligationmix:</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 300px;">
 +
<thead>
 +
<tr>
 +
<th scope="col" style="width: 79px;">Volume</th>
 +
<th scope="col" style="width: 213px;">Compound</th>
 +
</tr>
 +
</thead>
 +
<tbody>
 +
<tr>
 +
<td style="width: 67px;">10&nbsp;&mu;L</td>
 +
<td style="width: 201px;">pre-ligationmix&nbsp;</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">2&nbsp;&mu;L</td>
 +
<td style="width: 201px;">10x T4 ligationbuffer</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">1&nbsp;&mu;L</td>
 +
<td style="width: 201px;">T4 DNA ligase (1U/&mu;L)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">8&nbsp;&mu;L</td>
 +
<td style="width: 201px;">milli Q water</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 67px;">20&nbsp;&mu;L</td>
 +
<td style="width: 201px;">Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p>incubation at room-temperature for 20 minutes.<br>
 +
Store at -20 degrees Celcius</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>07</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 15</h2>
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</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">August 16</h3>
 +
<label class="back" for="switch-86">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>We've performed a colony PCR on 15 random (though not the red/purple -esque colonies, which are collored only because they contain BBa_J04450) colonies from J23105+B0032 on the 13th. We also ligated the following biobricks anew:<br>
 +
B0031 + J23105<br>
 +
B0031 + J23113<br>
 +
B0032 + J23100<br>
 +
B0032 + J23105<br>
 +
<br>
 +
We also transformed these biobricks into&nbsp;<em>E.coli</em>&nbsp;on the 16th, prepping and digesting them later on, though these and the PCR did not give us the result we'd hoped for.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Today a colony PCR will be performed to check our results from last friday, namely the transformation product of BBa_B0032 en BBa_J23105:<br>
 +
<br>
 +
Mastermix</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td><strong>What</strong></td>
 +
<td><strong>1 reaction</strong></td>
 +
<td><strong>30 reactions</strong></td>
 +
</tr>
 +
<tr>
 +
<td>Taq buffer(10x)</td>
 +
<td>5 ul</td>
 +
<td>150 ul</td>
 +
</tr>
 +
<tr>
 +
<td>10 mM dntp</td>
 +
<td>1 ul</td>
 +
<td>30 ul</td>
 +
</tr>
 +
<tr>
 +
<td>10 uM Forward primer</td>
 +
<td>1 ul</td>
 +
<td>30 ul</td>
 +
</tr>
 +
<tr>
 +
<td>10 uM Reverse primer</td>
 +
<td>1 ul</td>
 +
<td>30 ul</td>
 +
</tr>
 +
<tr>
 +
<td>Taq polymerase</td>
 +
<td>0,25 ul</td>
 +
<td>7,5 ul</td>
 +
</tr>
 +
<tr>
 +
<td>Nucease free hydrogen</td>
 +
<td>41,75 ul</td>
 +
<td>1252,5 ul</td>
 +
</tr>
 +
<tr>
 +
<td><strong>Total</strong></td>
 +
<td><strong>50 ul</strong></td>
 +
<td><strong>1500 ul</strong></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><br>
 +
15 colonies have been randomly selected to be checked via PCR.<br>
 +
<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>13</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h2 class="date" style="color: white;">August 16</h2>
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</svg>
 +
<h1 class="title">Testing gas production</h1>
 +
<h3 class="date">August 16</h3>
 +
<label class="back" for="switch-87">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>Making a set up for the gas production testing and testing it with NEB10B&egrave;ta with the expected biobricks in it (K133071 + J23100) and a negative control.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 14px;"><strong>Experiment 1: Gasproduction testing</strong></span><br>
 +
15-08-18<br>
 +
Making a set up for the gas production testing and testing it with NEB10B&egrave;ta with the expected biobricks in it (K133071 + J23100) and a negative control.<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
-15 mL tubes<br>
 +
-Little glass tubes<br>
 +
-Cocktail prickers<br>
 +
-Culture medium<br>
 +
-Urea<br>
 +
-Chloramphenicol (40 mg/mL)<br>
 +
<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Stick tape on the little tube and cover the little tubes with&nbsp;tin foil. This can now enter the autoclave.<br>
 +
Make overnight cultures (20 mL)of the desired colonies in culture medium with 17,5 ul/ 20mL Chloramphenicol.<br>
 +
<br>
 +
After a day, prepare the culture medium for the tests.<br>
 +
Put 1 g urea and 17,5 ul Chloramphenicol in 20 mL culture medium.<br>
 +
<br>
 +
Pipette&nbsp;10 mL of the overnight culture in the 15 mL tube en centrifuge&nbsp;5000 rpm fot 5 minutes. Throw the supernatant away.<br>
 +
Fill the 15 mL tube and the little tube to the edge with culture medium. Work sterile.<br>
 +
Put a paper on the little tube and turn it around.<br>
 +
Then hold it above the 15 mL tube and pull the paper away and let the little tube fall into the 15 mL tube.<br>
 +
Try to do this so there's no gas in the little tube.<br>
 +
Now wait until gas is produced.<br>
 +
<br>
 +
16-08-18<br>
 +
Making a set up for the gas production testing and testing it with NEB10B&egrave;ta with the expected biobricks in it (K133071 + J23100).<br>
 +
20 different colonies are tested.<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
Use the same materials as above.<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Use the protocol above.<br>
 +
Make 700 mL culture medium with 35&nbsp;g urea and 612,5 ul&nbsp;Chloramphenicol&nbsp;(40 mg/mL). This cannot be done sterile.<br>
 +
The culture medium can be used for the 20 different colonies with expected biobricks: K133071 + J23100.</p>
 +
 +
<p>&nbsp;<br>
 +
<strong>Results</strong><br>
 +
Gas was let into the little tubes so it was difficult to see whether there was gasproduction or not. Mainly in the negative control the tubes started with gas in it.<br>
 +
<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/a/aa/T--Rotterdam_HR--exp1F1.jpeg');"></div>
 +
  <p>Figure 1: Little tube with cocktailpricker and 15 mL tube</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/9/97/T--Rotterdam_HR--exp1F2.jpeg');"></div>
 +
  <p>Figure 2: Set up testing gas production</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/1/18/T--Rotterdam_HR--exp1F3.jpeg');"></div>
 +
  <p>Figure 3. tube 1,2,3 and 4 starting amounts of air</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/0/0d/T--Rotterdam_HR--Exp1F4.jpeg');"></div>
 +
  <p>Figure 4: After the weekend tube 1, 2, 3 and 4</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/6/65/T--Rotterdam_HR--exp1F5.jpeg');"></div>
 +
  <p>Figure 5. tube 5,6,7 and 8 starting amounts of air</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/e/e7/T--Rotterdam_HR--Exp1F6.jpeg');"></div>
 +
  <p>Figure 6: After the weekend tube 5, 6, 7 and 8</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/3/39/T--Rotterdam_HR--exp1F7.jpeg');"></div>
 +
  <p>Figure 7. tube 9 and 10 strating amounts of air</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/e/eb/T--Rotterdam_HR--exp1F8.jpeg');"></div>
 +
  <p>Figure 8: After the weekend tube 9 and 10</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/2/23/T--Rotterdam_HR--exp1F9.jpeg');"></div>
 +
  <p>Figure 9. tube 11,12,13,and 14 starting amounts of air</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/7/7f/T--Rotterdam_HR--exp1F10.jpeg');"></div>
 +
  <p>Figure 10: After the weekend tube 11, 12, 13 and 14</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/5/5f/T--Rotterdam_HR--exp1F11.jpeg');"></div>
 +
  <p>Figure 11. tube 15,16,17 and 18 starting amounts of air</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/f6/T--Rotterdam_HR--exp1F12.jpeg');"></div>
 +
  <p>Figure 12: After the weekend tube 15, 16, 17 and 18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/b/bc/T--Rotterdam_HR--exp1F13.jpeg');"></div>
 +
  <p>Figure 13. tube 19 and 20 starting amounts of air</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/b/b1/T--Rotterdam_HR--exp1F14.jpeg');"></div>
 +
  <p>Figure 14: After the weekend tube 19 and 20</p>
 +
</div>
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing gas production</h3>
 +
<svg viewBox="0 0 24 24">
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<h2 class="date" style="color: white;">August 16</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 21</h3>
 +
<label class="back" for="switch-88">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Minipreps are made of colonies 11 and 19 of biobrick combination&nbsp;K133071 + J23100.&nbsp;Results: 11. 270,57 ng/ul&nbsp;19. 253,59 ng/ul.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Minipreps</strong></span><br>
 +
21-08-18<br>
 +
<br>
 +
Minipreps of colonies 11 and 19 (see Testing gasproduction experiment 1)<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
- Mini prep, Plasmid Purification Kit, Machery Michels&nbsp;<br>
 +
- Eppendorf cups<br>
 +
- Variable Volume Pipets<br>
 +
- sterile pipet points<br>
 +
-Chloramphenicol (40 mg/mL)<br>
 +
<br>
 +
<strong>Methods&nbsp;</strong><br>
 +
<strong>Miniprep protocol&nbsp;Plasmid Purification Kit, Machery Nagel</strong><br>
 +
See experiment 3<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
<br>
 +
11. K133071 + J23100: 270,57 ng/ul<br>
 +
19. K133071 + J23100: 253,59 ng/ul</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/3/3c/T--Rotterdam_HR--Nanodrop21_08.jpeg');"></div>
 +
<p>Figure 1: Results minipreps colonies 11 and 19</p>
 +
  </div>  
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>08</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<h2 class="date" style="color: white;">August 21</h2>
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<h1 class="title">Testing gas production</h1>
 +
<h3 class="date">August 22</h3>
 +
<label class="back" for="switch-89">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>Making a set up for the gas production testing and testing it with NEB10B&egrave;ta with the expected biobricks in it (K133071 + I13453), (K173003 + J23100), (K173003 + I13453)&nbsp;and a negative control.&nbsp;Colonies 9 and 10 are chosen use for further tests because there was the biggest amount of gas production.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 14px;"><strong>Experiment 2: Gasproduction testing </strong></span><br>
 +
<strong>20-08-18</strong><br>
 +
<br>
 +
Making a set up for the gas production testing and testing it with NEB10B&egrave;ta with the expected biobricks in it (K133071 + I13453), (K173003 + J23100), (K173003 + I13453)&nbsp;and a negative control<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
See materials Experiment 1<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See methods Experiment 1<br>
 +
850 mL culture medium is made with 21,5 g LB for 42 erlenmeyers (also for CooA production).<br>
 +
Overnight cultures are made of 10 colonies each of K173003&nbsp;+ I13453 and K173003 + J23100.<br>
 +
<br>
 +
<strong>21-08-18</strong><br>
 +
<br>
 +
K173003 + J23100 are tested (see methods experiment 1)<br>
 +
Overnight cultures are made of&nbsp; K133071 + I13453 and K173003 + I13453 are tested later.<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
Colonies 9 and 10 are chosen use for further tests.</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/fd/T--Rotterdam_HR--exp2F1.jpeg');"></div>
 +
  <p>Figure 1. Starting amounts of air in tube 1,2,3 and 4.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/8/8c/T--Rotterdam_HR--exp2F2.jpeg');"></div>
 +
  <p>Figure 2. Gas formation in tube 1,2,3 and 4. 30 minutes after adding
 +
Sodium pyruvate.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/6/63/T--Rotterdam_HR--exp2f3.jpeg');"></div>
 +
  <p>Figure 3. Starting amounts of air in tube 5,6,7 and 8.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/3/34/T--Rotterdam_HR--exp2f4.jpeg');"></div>
 +
  <p>Figure 4. Gas formation in tube 5,6,7 and 8. 30 minutes after adding Sodium pyruvate.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/e/e6/T--Rotterdam_HR--exp2f5.jpeg');"></div>
 +
  <p>Figure 5. Starting amounts of air in tube 9 and 10.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/0/0d/T--Rotterdam_HR--exp2f6.jpeg');"></div>
 +
  <p>Figure 6. Gas formation in tube 9 and 10. 30 minutes after adding
 +
Sodium pyruvate.
 +
</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/9/96/T--Rotterdam_HR--exp2f7.jpeg');"></div>
 +
  <p>Figure 7: Gas formation in tube 1, 2, 3 and 4 after a dayr</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/7/70/T--Rotterdam_HR--exp2f8.jpeg');"></div>
 +
  <p>Figure 8: Gas formation in tube 5, 6, 7 and 8 after a day</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/6/6f/T--Rotterdam_HR--exp2f9.jpeg');"></div>
 +
  <p>Figure 9: Gas formation in tube 9 and 10 after a day</p>
 +
</div>
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing gas production</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 22</h2>
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 +
<h1 class="title">Testing gas production</h1>
 +
<h3 class="date">August 22</h3>
 +
<label class="back" for="switch-90">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>Gasproduction testing for the biobricks (K173003 + I13453), (K133071 + I13453) and negative controls (B0015 and K133071 without urea and arabinose).<br>
 +
K173003 + I13453 is tested with sodiumpyruvate and arabinose for gasproduction and&nbsp;K133071 + I13453&nbsp;is tested with urea and arabinose for gasproduction.<br>
 +
The negative control also produces a little bit gas.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 3: Gasproduction testing</strong></span><br>
 +
<br>
 +
Gasproduction testing for the biobricks K173003 + I13453,&nbsp;K133071 + I13453 and negative controls.<br>
 +
K173003 + I13453 is tested with sodiumpyruvate and arabinose for gasproduction and&nbsp;K133071 + I13453&nbsp;is tested with urea and arabinose for gasproduction.<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
See experiment 1 and 2.<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See experiment 1 and 2.<br>
 +
375 mL culture medium with CAM (already in it) + 18,75&nbsp;g urea + 0,5 ul arabinose.<br>
 +
340 mL culture medium with CAM (already in it) + 51 mL natriumpyruvate (20 mM) + 0,5 ul arabinose.<br>
 +
<br>
 +
<br>
 +
<strong>Results and conclusion</strong><br>
 +
Colonies 5 and 7 of the biobricks K173003 + I13453 (pyruvate)&nbsp; and number 4 of the biobricks K133071 + I13453 (urea).<br>
 +
In both the negative controls there's been gas produced. Futher gasproduction test will be done.</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/2/24/T--Rotterdam_HR--exp3Fi1.jpeg');"></div>
 +
  <p>Figure 1: Starting amounts in tube 1, 2, 3, 4 and 5 with Pyruvate and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/8/8c/T--Rotterdam_HR--exp3f2.jpeg');"></div>
 +
  <p>Figure 2: Gasproduction in tube 1, 2, 3, 4 and 5 after a day with pyruvaat and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/c/c0/T--Rotterdam_HR--exp3f3.jpeg');"></div>
 +
  <p>Figure 3: Starting amounts in tube 6, 7, 8, 9 and 10 with Pyruvate and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/b/bc/T--Rotterdam_HR--exp3f4.jpeg');"></div>
 +
  <p>Figure 4: Gasproduction in tube 6, 7, 8, 9 and 10 after a day with pyruvate and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/ff/T--Rotterdam_HR--exp3f5.jpeg');"></div>
 +
  <p>Figure 5: Starting amounts in tube 1, 2, 3, 4 and 5 with Urea and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/d/d3/T--Rotterdam_HR--exp3f6.jpeg');"></div>
 +
  <p>Figure 6: Starting amounts in tube 6, 7, 8, 9 and 10 with Urea and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/a/ac/T--Rotterdam_HR--exp3F7.jpeg');"></div>
 +
  <p>Figure 7: Gasproduction in tube 1, 2, 3, 4 and 5 after a day with urea and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/a/ab/T--Rotterdam_HR--exp3f8.jpeg');"></div>
 +
  <p>Figure 8: Gasproduction in tube 6, 7, 8, 9 and 10 after a day with urea and arabinose</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/b/b7/T--Rotterdam_HR--exp3f9.jpeg');"></div>
 +
  <p>Figure 9: Starting amounts in the negative controls B0015 and K133071 + I13453 (without urea and arabinose)</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/9/9a/T--Rotterdam_HR--exp3f10.jpeg');"></div>
 +
  <p>Figure 10: Gasproduction in the negative controls B0015 and K133071 + I13453 after a day (without urea and arabinose)</p>
 +
</div>
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing gas production</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 22</h2>
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<h1 class="title">Testing gas production</h1>
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<h3 class="date">August 24</h3>
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<label class="back" for="switch-91">&times;</label>
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<div class="attendees">Elise Grootscholten | Paul Reusink</div>
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<div class="description">
 +
<p>Testing the gasproduction&nbsp;of the colonies&nbsp;9 and 10 of biobricks (K173003 + J23100), colonies 5 and 7 of biobricks (K173003 + I13453) and colony 4 of&nbsp;biobricks (K133071 + I13453) with and without centrifuging the bacteria. There is also a negative control&nbsp;(J04450 pSB1K3) with Kanamycine. The negative contol started with a lot of gas inside the tube. We can not see wether there is produced more after a day or not. This have to be tested later.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 4: Gasproduction testing&nbsp;</strong></span><br>
 +
24-08-18<br>
 +
<br>
 +
Testing the gasproduction with and without centrifuging the bacteria.<br>
 +
<br>
 +
<strong>methods</strong><br>
 +
See methods experiment 1<br>
 +
2 negative controls<br>
 +
colonies 9 and 10 of biobricks&nbsp;K173003 + J23100.<br>
 +
Colonies 5 and 7 of biobricks&nbsp;K173003 + I13453<br>
 +
Colony 4 of&nbsp;biobricks K133071 + I13453<br>
 +
<br>
 +
Culture medium:<br>
 +
150 mL: Sodiumpyruvate (20 mM) 22,5 mL +&nbsp;Chloramphenicol (40 mg/mL) 131 ul<br>
 +
150 mL: Sodiumpyruvate (20mM) 22,5 mL +&nbsp;&nbsp;Chloramphenicol (40 mg/mL) 131 ul + Arabinose (300 mg/mL) 0,5 ul<br>
 +
80 mL: Urea 4 g +&nbsp;Chloramphenicol (40 mg/mL) 70 ul +&nbsp;Arabinose (300 mg/mL) 0,5 ul<br>
 +
80 mL: Kanamycine (40 mg/mL) 70 ul<br>
 +
<br>
 +
&nbsp;</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/e/e7/T--Rotterdam_HR--exp4f1.jpeg');"></div>
 +
  <p>Figure 1: Starting amounts Nr. 9, 10 and 4 centrifuged </p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/8/83/T--Rotterdam_HR--exp4f2.jpeg');"></div>
 +
  <p>Figure 2. Gasproduction Nr 9. 10 and 4 after 4 hours centrifuged.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/a/a1/T--Rotterdam_HR--exp4f3.jpeg');"></div>
 +
  <p>Figure 3: Starting amounts Nr. 5, 7 and negative control centrifuged</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/f2/T--Rotterdam_HR--exp4f4.jpeg');"></div>
 +
  <p>Figure 4. Gasproduction Nr 5,7 and negative control after 4 hours centrifuged.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/d/df/T--Rotterdam_HR--exp4f5.jpeg');"></div>
 +
  <p>Figure 5: Starting amounts Nr. 9, 10 and 4 not  centrifuged </p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/a/a8/T--Rotterdam_HR--exp4f6.jpeg');"></div>
 +
  <p>Figure 6. Gasproduction Nr 9. 10 and 4 after 4 hours not centrifuged.</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/8/88/T--Rotterdam_HR--exp4f7.jpeg');"></div>
 +
  <p>Figure 7: Starting amounts Nr. 5, 7 and negative control not centrifuged</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/1/15/T--Rotterdam_HR--exp4f8.jpeg');"></div>
 +
  <p>Figure 8. Gasproduction Nr. 5, 7 and negative control after 4 hours, not centrifuged.</p>
 +
</div>
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing gas production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<label for="switch-91" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">August 24</h2>
 +
</div>
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 +
</div><style>
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Urea and sodium pyruvate test for resistance E.coli</h1>
 +
<h3 class="date">August 28</h3>
 +
<label class="back" for="switch-92">&times;</label>
 +
</div>
 +
<div class="attendees">Randall de Waard | Elise Grootscholten</div>
 +
<div class="description">
 +
<p><span style="font-size:14px;">Testing different amounts of urea and sodiumpyruvate to know which concentrations the bacteria survive.</span></p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size: 16px;"><strong>Experiment 2:&nbsp;</strong></span><strong><span style="font-size:16px;">Testing different concentrations of Urea and Sodium pyruvate</span></strong><br>
 +
<strong><span style="font-size:16px;">13-08-18</span></strong><br>
 +
<br>
 +
<br>
 +
<span style="font-size:14px;">Same testing like experiment 1 but with different concentrations.</span></p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 420px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 163px;"><span style="font-size: 14px;"><strong>What</strong></span></td>
 +
<td style="width: 84px;"><span style="font-size: 14px;"><strong>How much</strong></span></td>
 +
<td style="width: 157px;"><span style="font-size: 14px;"><strong>Solution</strong></span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;"><span style="font-size: 14px;">Urea</span></td>
 +
<td style="width: 84px;"><span style="font-size: 14px;">1,2 g</span></td>
 +
<td style="width: 157px;"><span style="font-size: 14px;">1 mL purified water</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;"><span style="font-size: 14px;">Urea</span></td>
 +
<td style="width: 84px;"><span style="font-size: 14px;">1,6 g</span></td>
 +
<td style="width: 157px;"><span style="font-size: 14px;">2 mL purified water</span></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 163px;"><span style="font-size: 14px;">Sodium pyruvate (10 mM)</span></td>
 +
<td style="width: 84px;"><span style="font-size: 14px;">10 mL</span></td>
 +
<td style="width: 157px;"><span style="font-size: 14px;">None</span></td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<br>
 +
<br>
 +
<span style="font-size:14px;"><strong>Results</strong></span><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 564px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 198px;"><span style="font-size:14px;"><strong>What</strong></span></td>
 +
<td style="width: 93px;"><span style="font-size:14px;"><strong>How much</strong></span></td>
 +
<td style="width: 158px;"><span style="font-size:14px;"><strong>Solution</strong></span></td>
 +
<td style="width: 93px;"><strong><span style="font-size:14px;">Result</span></strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 198px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 93px;"><span style="font-size:14px;">1,2 g</span></td>
 +
<td style="width: 158px;"><span style="font-size:14px;">1 mL purified water</span></td>
 +
<td style="width: 93px;">Little grow</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 198px;"><span style="font-size:14px;">Urea</span></td>
 +
<td style="width: 93px;"><span style="font-size:14px;">1,6 g</span></td>
 +
<td style="width: 158px;"><span style="font-size:14px;">2 mL purified water</span></td>
 +
<td style="width: 93px;">No grow</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 198px;"><span style="font-size:14px;">Sodium pyruvate (10 mM)</span></td>
 +
<td style="width: 93px;"><span style="font-size:14px;">10 mL</span></td>
 +
<td style="width: 158px;"><span style="font-size: 14px;">-</span></td>
 +
<td style="width: 93px;">Grow</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 198px;"><span style="font-size:14px;">Positive control</span></td>
 +
<td style="width: 93px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 158px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 93px;">Grow</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 198px;"><span style="font-size:14px;">Negative control</span></td>
 +
<td style="width: 93px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 158px;"><span style="font-size:14px;">-</span></td>
 +
<td style="width: 93px;">No Grow</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 2: Results experiment 2</em><br>
 +
<br>
 +
<span style="font-size:14px;"><strong>Conclusion</strong><br>
 +
<br>
 +
For urea is 1 g used/20 mL and for sodiumpyruvate is 6 mL (10mM) or 3 mL (20 mM) used.</span><br>
 +
&nbsp;
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Urea and sodium pyruvate test for resistance E.coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
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<label for="switch-92" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">August 28</h2>
 +
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 +
</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 30</h3>
 +
<label class="back" for="switch-93">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>A miniprep of the biobrick K352002 is made. The concentration is 69,77 ng/ul.</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Experiment 9: Minipreps</strong><br>
 +
28-08-18<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
- Mini prep, Plasmid Purification Kit, Machery Nagel<br>
 +
- Eppendorf cups<br>
 +
- Variable Volume Pipets<br>
 +
- sterile pipet points<br>
 +
-Chloramphenicol (40 mg/mL)<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
<strong>Miniprep protocol&nbsp;Plasmid Purification Kit, Machery Nagel</strong><br>
 +
See experiment 3<br>
 +
Make minipreps of the biobricks K352002, K352003<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
The minipreps can not be used,&nbsp;because washing buffer without ethanol is used. The miniprep of K352002&nbsp;has&nbsp;been done again.<br>
 +
<br>
 +
29-08-18<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Make a miniprep&nbsp;of the biobrick&nbsp;K352002<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
K352002: 69,77 ng/&mu;L<br>
 +
<br>
 +
30-08-18<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See experiment 3<br>
 +
Make minipreps of the biobricks K133116 and K173013</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/3/32/T--Rotterdam_HR--Nanodrop29_08.jpeg');"></div>
 +
<p>Figure 1: Miniprep K352002</p>
 +
  </div>  
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>09</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 30</h2>
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 +
</svg>
 +
<h1 class="title">Chemo competent cells </h1>
 +
<h3 class="date">August 30</h3>
 +
<label class="back" for="switch-94">&times;</label>
 +
</div>
 +
<div class="attendees">None</div>
 +
<div class="description">
 +
<p>Competent cells of NEB10 Beta and BL21 DE3 were made. These cells were made competent so new biobricks and ligation mixes could be transformed into these cells. BL21 DE3 and NEB10 beta had 80 cups of 50 ul each. 1 cup is needed for 1 transformation later. The cells were competent enough to use in the project.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p>Making chemocompetent cells:&nbsp;<br>
 +
<br>
 +
Materials:&nbsp;<br>
 +
- E. coli NEB10Beta strain&nbsp;<br>
 +
- E. coli Bl21 (DE 3) strain<br>
 +
- Buffer 1&nbsp;</p>
 +
 +
<p>30 mM natriumacetaat, 100mM rubidium chloride,10 mM calciumchloride, 50mM mangaan chloride, 15 % glycerol; pH 5,8 met verdunde azijnzuur, filter steriliseren.</p>
 +
 +
<p>Bewaren bij kamertemperatuur.</p>
 +
 +
<p>- Buffer 2&nbsp;</p>
 +
 +
<p>10 mM MOPS, 75 mM calcium chloride, 10 mM rubidium chloride, 15% glycerol; pH 6,6 met NaOH, filter steriliseren.</p>
 +
 +
<p>Bewaren bij kamertemperatuur&nbsp;<br>
 +
<br>
 +
Method:&nbsp;<br>
 +
The day before making competent cells the strains were ented into 20ml LB medium (100ml erlenmeyer)&nbsp;and incubated for 6 hours by 37 degrees at 200 rpm. The culture was then transmitted to 20 ml LB medium (100 ml erlenmeyer)&nbsp;in a dilution of 1:50&nbsp;(400 ul/ 20 ML) this was incubated overnight at 37 degrees Celsius at 200 rpm.&nbsp;<br>
 +
<br>
 +
In the morning the culture was diluted 1:100 in 200 ml LB medium (1L erlenmeyer). This was then incubated by 37 degrees and 225 rpm till a OD600 level of 0,5.&nbsp;<br>
 +
To make sure the right OD was used, the culture was measured every 20/30 minutes. The results of the measurements can be found in part 2.&nbsp;<br>
 +
<br>
 +
After hitting the OD600 of 0,5. The cultures were&nbsp;transmitted to 250 ml centrifuge tubes and cooled on ice. The cells are supposed to stay on ice as much as possible&nbsp;after this.&nbsp;After cooling down to 4 degrees, the cultures&nbsp;were centrifuged at 4500 rpm for 10 min at 4 degrees. The supernatant were disposed, the pellet was resuspended in 66,4 ml ice cold Buffer 1(see materials). This was centrifuged &nbsp;at 5000 rpm for 10 min at 4 degrees.&nbsp;<br>
 +
<br>
 +
<br>
 +
BL21DE3 &nbsp;80 cups on place: Drawer 3, tower 2, drawer 3, place 2.&nbsp;<br>
 +
NEB10Beta 79 cups on place: Drawer 3, tower 2, drawer 3, place 3.&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Chemo competent cells </h3>
 +
<svg viewBox="0 0 24 24">
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<h2 class="date" style="color: white;">August 30</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">August 31</h3>
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<label class="back" for="switch-95">&times;</label>
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</div>
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<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Digestions, gelelectrophoresis, dephosphorylations and ligations of different biobrick combinations. These biobrick combinations are 4 different promoters (J23100, I13453, K352002, K352003)&nbsp; with 4 different gasproduction biobricks (k173003, K173013, K133071, K133116). J23100 is about 1 kb to long. The rest seems likely to be right digested. The ligations will be transformed in NEB10b&egrave;ta and digested again as control.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 10: Digestion, defosforylation and ligation</strong></span><br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
-Cutsmart buffer (10x)<br>
 +
-Restriction enzymes XbaI, PstI-HF, EcoRI-HF, SpeI<br>
 +
-Nuclease free water<br>
 +
-DNA<br>
 +
-Phosphatase buffer (10x)<br>
 +
-Phosphatase (1 U/&mu;L)<br>
 +
-Delutionbuffer (5x)</p>
 +
 +
<p>-T4 ligationbuffer&nbsp;(10x)<br>
 +
-T4 DNA ligase (1U/&mu;L)<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (J23100 325,59 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">6,1</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><em>Table 1: Digestion scheme 1</em><br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (J23100 325,59 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">6,1</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 2: Digestion scheme 2</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (J23100 325,59 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">6,1</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 3: Digestion scheme 3</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 351px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 222px;">DNA (K352003 180,65 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 117px;">11,1&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Cutsmart (10x)</td>
 +
<td style="width: 117px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 117px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 117px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Nuclease free water</td>
 +
<td style="width: 117px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 4: Digestion scheme 4</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 351px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 222px;">DNA (K352002 69,77 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 117px;">28,7 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Cutsmart (10x)</td>
 +
<td style="width: 117px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 117px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 117px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Nuclease free water</td>
 +
<td style="width: 117px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 5: Digestion scheme 5</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 352px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 224px;">DNA (K173003 217,06 ng/&mu;L)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 116px;">9,2&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 224px;">Cutsmart (10x)</td>
 +
<td style="width: 116px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 224px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 116px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 224px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 116px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 224px;">Nuclease free water</td>
 +
<td style="width: 116px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 6: Digestion scheme 6</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 359px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 227px;">DNA (I13453 88,18 ng/&mu;L)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 121px;">22,7&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 227px;">Cutsmart (10x)</td>
 +
<td style="width: 121px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 227px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 121px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 227px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 121px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 227px;">Nuclease free water</td>
 +
<td style="width: 121px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 7: Digestion scheme 7</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 362px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 228px;">DNA (K133071 186,79&nbsp;ng/&mu;L)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 122px;">10,7&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Cutsmart (10x)</td>
 +
<td style="width: 122px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 122px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 122px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Nuclease free water</td>
 +
<td style="width: 122px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 8: Digestion scheme 8</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 358px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 228px;">DNA (pSB1K3 163,93 ng/&mu;L)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 118px;">&nbsp;12,2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Cutsmart (10x)</td>
 +
<td style="width: 118px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 118px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 118px;">2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 228px;">Nuclease free water</td>
 +
<td style="width: 118px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 9: Digestion scheme 9</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 357px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 222px;">DNA (K173013 345,1 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 123px;">&nbsp;5,8 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Cutsmart (10x)</td>
 +
<td style="width: 123px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 123px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 123px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Nuclease free water</td>
 +
<td style="width: 123px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 10: Digestion scheme 10</em>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 357px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 222px;">DNA (K133116 194,1 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 123px;">10,3&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Cutsmart (10x)</td>
 +
<td style="width: 123px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 1 (1 U/&mu;L)</td>
 +
<td style="width: 123px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Restriction enzyme 2 (1 U/&mu;L)</td>
 +
<td style="width: 123px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 222px;">Nuclease free water</td>
 +
<td style="width: 123px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 11: Digestion scheme 11</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 471px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 124px;"><strong>Biobrick</strong></td>
 +
<td style="width: 158px;"><strong>Restriction-enzyme 1</strong></td>
 +
<td style="width: 172px;"><strong>Restriction enzyme 2</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">K173003</td>
 +
<td style="width: 158px;">XbaI</td>
 +
<td style="width: 172px;">PstI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">K173013</td>
 +
<td style="width: 158px;">XbaI</td>
 +
<td style="width: 172px;">PstI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">K133071</td>
 +
<td style="width: 158px;">XbaI</td>
 +
<td style="width: 172px;">PstI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">K133116</td>
 +
<td style="width: 158px;">XbaI</td>
 +
<td style="width: 172px;">PstI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">J23100</td>
 +
<td style="width: 158px;">EcoRI-HF</td>
 +
<td style="width: 172px;">SpeI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">I13453</td>
 +
<td style="width: 158px;">EcoRI-HF</td>
 +
<td style="width: 172px;">SpeI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">K352002</td>
 +
<td style="width: 158px;">EcoRI-HF</td>
 +
<td style="width: 172px;">SpeI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">K352003</td>
 +
<td style="width: 158px;">EcoRI-HF</td>
 +
<td style="width: 172px;">SpeI-HF</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 124px;">pSB1K3</td>
 +
<td style="width: 158px;">EcoRI-HF</td>
 +
<td style="width: 172px;">PstI-HF</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 12: Digestion scheme used enzymes</em><br>
 +
<br>
 +
<strong>Gelelectrophoresis</strong>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 357px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 69px;"><strong>Number</strong></td>
 +
<td style="width: 170px;"><strong>What</strong></td>
 +
<td style="width: 101px;"><strong>Amount</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">1.</td>
 +
<td style="width: 170px;">Ladder</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">2.</td>
 +
<td style="width: 170px;">J23100 EcoRI-HF+ SPeI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">3.</td>
 +
<td style="width: 170px;">J23100 EcoRI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">4.</td>
 +
<td style="width: 170px;">J23100 SpeI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">5.</td>
 +
<td style="width: 170px;">pSB1K3 EcoRI-HF + PstI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">6</td>
 +
<td style="width: 170px;">K173013 XbaI + PstI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">7.</td>
 +
<td style="width: 170px;">K173003 XbaI + PstI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">8.</td>
 +
<td style="width: 170px;">K133071 XbaI + PstI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">9.</td>
 +
<td style="width: 170px;">K133116 XbaI + PstI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">10.</td>
 +
<td style="width: 170px;">K352002 EcoRI-HF + SpeI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">11.</td>
 +
<td style="width: 170px;">K352003 EcoRI-HF + SpeI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">12.</td>
 +
<td style="width: 170px;">I13453 EcoRI-HF + SpeI-HF</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">13.</td>
 +
<td style="width: 170px;">Ladder&nbsp;</td>
 +
<td style="width: 101px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 69px;">14.</td>
 +
<td style="width: 170px;">Ladder</td>
 +
<td style="width: 101px;">10 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 13: Gelelectrophoresis order of digestions</em><br>
 +
<br>
 +
<strong>dephosphorylation</strong>
 +
 +
<p>&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 219px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 58px;"><strong>Volume</strong></td>
 +
<td style="width: 149px;"><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">12,5 &mu;L</td>
 +
<td style="width: 149px;">Vector DNA (0,5&nbsp;&mu;g)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">2 &mu;L</td>
 +
<td style="width: 149px;">phosphatase buffer (10x)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">4,5 &mu;L</td>
 +
<td style="width: 149px;">milli Q</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">1&nbsp;&mu;L</td>
 +
<td style="width: 149px;">phosphatase (1 U/&mu;L)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">20 &mu;L</td>
 +
<td style="width: 149px;">Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 14: Pipetting scheme&nbsp;dephosphorylation</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 219px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 58px;"><strong>Volume</strong></td>
 +
<td style="width: 149px;"><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">37,5 &mu;L</td>
 +
<td style="width: 149px;">Vector DNA (0,5&nbsp;&mu;g)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">6 &mu;L</td>
 +
<td style="width: 149px;">phosphatase buffer (10x)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">13,5 &mu;L</td>
 +
<td style="width: 149px;">milli Q</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">3&mu;L</td>
 +
<td style="width: 149px;">phosphatase (1 U/&mu;L)</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 58px;">60 &mu;L</td>
 +
<td style="width: 149px;">Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 15: Pipetting scheme dephosphorylation</em><br>
 +
<br>
 +
<strong>Pre-Ligationmix:</strong>
 +
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td><strong>Volume</strong></td>
 +
<td><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td>7,5 &mu;L</td>
 +
<td>Insert DNA (2 &mu;g/50 &mu;L) 300 ng</td>
 +
</tr>
 +
<tr>
 +
<td>7,5 &mu;L</td>
 +
<td><font face="arial, helvetica, sans-serif">Insert DNA (2&nbsp;</font>&mu;g/50 &mu;L) 300 ng</td>
 +
</tr>
 +
<tr>
 +
<td>4&nbsp;&mu;L</td>
 +
<td>Backbone (0,5&nbsp;&mu;g/ 20&nbsp;&mu;L) 100 ng</td>
 +
</tr>
 +
<tr>
 +
<td>5 &mu;L</td>
 +
<td>Delutionbuffer (5x)</td>
 +
</tr>
 +
<tr>
 +
<td>1 &mu;L</td>
 +
<td>Milli Q</td>
 +
</tr>
 +
<tr>
 +
<td>25 &mu;L</td>
 +
<td>Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 16: Pre-ligationmix pipette scheme</em><br>
 +
<br>
 +
<strong>Ligationmix:</strong>
 +
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td><strong>Volume</strong></td>
 +
<td><strong>Compound</strong></td>
 +
</tr>
 +
<tr>
 +
<td>10 &mu;L</td>
 +
<td>Pre-ligationmix</td>
 +
</tr>
 +
<tr>
 +
<td>2 &mu;L</td>
 +
<td>10x T4 ligationbuffer</td>
 +
</tr>
 +
<tr>
 +
<td>1 &mu;L</td>
 +
<td>T4 DNA ligase (1U/&mu;L)</td>
 +
</tr>
 +
<tr>
 +
<td>8 &mu;L</td>
 +
<td>Milli Q</td>
 +
</tr>
 +
<tr>
 +
<td>20&nbsp;&mu;L</td>
 +
<td>Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 17: Ligationmix pipette scheme</em><br>
 +
<br>
 +
Ligation started at 15.45 (31-08-18) at room temperature.<br>
 +
end ligation around 11.00 (01-09-18)<br>
 +
Store at -20 degrees Celcius.<br>
 +
&nbsp;
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>10</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">August 31</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 3</h3>
 +
<label class="back" for="switch-96">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Transformations of the biobrick&nbsp;combinations in NEB10b&egrave;ta. The transformations are plated on kanamycine agar plates, because all the ligations were done in pSB1K3 (kanamycine resistence) backbone. Pink and with colonies appeared after incubation by 37 degrees Celcius for about 12h. The white colonies will be used for further experiments.</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Experiment 11: transformations of the biobrick ombinations in NEB10b&egrave;ta.</strong><br>
 +
03-09-18<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
See materials&nbsp;experiment 5.<br>
 +
-Kanamycin&nbsp;(40 mg/mL)<br>
 +
-Ampicillin (50 mg/mL)<br>
 +
<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See methods experiment 5<br>
 +
<br>
 +
Make 19 agar plates with kanamycine (Kana) (17,5 ul/20mL) and 1 agar plate with ampicillin (Amp) (40 ul/mL). Plate all the transformations on agar plates with Kana , because all the ligations are done in the pSB1K3 (Kana resistence) backbone.&nbsp;<br>
 +
Devide one Amp plate and one Kana plate in about 30 parts. Because the biobrick combinations with biobrick K133116 have originally Amp and Kana resistence, the right backbone plasmid (with hopefully the right ligation) is only growing on the agar plate with Kana.<br>
 +
<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
There is grown n every transformation plates, so the transformations are done well.<br>
 +
There are pink and white colonies present. Only the white colonies will be used for further experiments.<br>
 +
&nbsp;</p>
 +
 +
</div>
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<div class="cardform">
 +
<div class="number"><b>11</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<h2 class="date" style="color: white;">September 3</h2>
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<h1 class="title">Soldering PCB [TC]</h1>
 +
<h3 class="date">September 4</h3>
 +
<label class="back" for="switch-97">&times;</label>
 +
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 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Soldered the first temperature controller with the help of a sodlering stencil and reflow oven. All components were soldered onto the first temperature controller.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
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 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Soldering PCB [TC]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Soldering PCB [TC]</h1>
 +
<h3 class="date">September 4</h3>
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<label class="back" for="switch-98">&times;</label>
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</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Soldered second temperature controller with the help of a sodlering stencil and reflow oven. All components were soldered onto the first temperature controller.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>02</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Soldering PCB [TC]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-98" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 4</h2>
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</div>
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</div><style>
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 6</h3>
 +
<label class="back" for="switch-99">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Plasmid DNA was isolated of 20 different colonies. The first time something went wrong. The second time we had good concentrations of isolated plasmid DNA. After this we will digest the DNA to see if the plasmids all have the required biobricks. The tube have a code from now on, see table 1.<br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 122px;"><strong>Number</strong></td>
 +
<td style="width: 133px;"><strong>Promotor</strong></td>
 +
<td style="width: 116px;"><strong>Gene</strong></td>
 +
<td style="width: 106px;"><strong>Backbone</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">A</td>
 +
<td style="width: 133px;">K352002 (CooF)</td>
 +
<td style="width: 116px;">K173013</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">B</td>
 +
<td style="width: 133px;">K352002 (CooF)</td>
 +
<td style="width: 116px;">K173003</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">C</td>
 +
<td style="width: 133px;">K352002 (CooF)</td>
 +
<td style="width: 116px;">K133071</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">D</td>
 +
<td style="width: 133px;">K352003 (CooM)</td>
 +
<td style="width: 116px;">K133071</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">E</td>
 +
<td style="width: 133px;">K352003 (CooM)</td>
 +
<td style="width: 116px;">K173003</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">F</td>
 +
<td style="width: 133px;">K352003 (CooM)</td>
 +
<td style="width: 116px;">K173013</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">H</td>
 +
<td style="width: 133px;">I13453</td>
 +
<td style="width: 116px;">K173013</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">I</td>
 +
<td style="width: 133px;">I13453</td>
 +
<td style="width: 116px;">K173003</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">J</td>
 +
<td style="width: 133px;">I13453</td>
 +
<td style="width: 116px;">K133071</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><em>Table 1: list of biobricks of abbreviations</em></p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Experiment&nbsp;12: Overnight cultures and minipreps</strong><br>
 +
05-09-18/06-09-18<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
See materials experiment 3<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See methods experiment 3</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 122px;"><strong>Number</strong></td>
 +
<td style="width: 131px;"><strong>Promotor</strong></td>
 +
<td style="width: 118px;"><strong>Gene</strong></td>
 +
<td style="width: 106px;"><strong>Backbone</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">A</td>
 +
<td style="width: 131px;">K352002 (CooF)</td>
 +
<td style="width: 118px;">K173013</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">B</td>
 +
<td style="width: 131px;">K352002 (CooF)</td>
 +
<td style="width: 118px;">K173003</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">C</td>
 +
<td style="width: 131px;">K352002 (CooF)</td>
 +
<td style="width: 118px;">K133071</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">D</td>
 +
<td style="width: 131px;">K352003 (CooM)</td>
 +
<td style="width: 118px;">K133071</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">E</td>
 +
<td style="width: 131px;">K352003 (CooM)</td>
 +
<td style="width: 118px;">K173003</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">F</td>
 +
<td style="width: 131px;">K352003 (CooM)</td>
 +
<td style="width: 118px;">K173013</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">G</td>
 +
<td style="width: 131px;">B0032</td>
 +
<td style="width: 118px;">J23105</td>
 +
<td style="width: 106px;">pSB1A3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">H</td>
 +
<td style="width: 131px;">I13453</td>
 +
<td style="width: 118px;">K173013</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">I</td>
 +
<td style="width: 131px;">I13453</td>
 +
<td style="width: 118px;">K173003</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 122px;">J</td>
 +
<td style="width: 131px;">I13453</td>
 +
<td style="width: 118px;">K133071</td>
 +
<td style="width: 106px;">pSB1K3</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
Table 1: list of biobricks of abbreviations<br>
 +
&nbsp;
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/8/80/T--Rotterdam_HR--Nanodrop06_09.jpeg');"></div>
 +
<p>Figure 1: Nanodrop results after DNA isolation. The biobrick combinations can be found in the methods (experiment 12)</p>
 +
  </div>  
 +
</div>
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</div>
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 +
<div class="cardform">
 +
<div class="number"><b>12</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">September 6</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">September 6</h3>
 +
<label class="back" for="switch-100">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
After transforming the ligation product of J23105+B0032 and K352001+B0015 we had some growth of none red/pink colonies. Overnight cultures where made, and the DNA was prepped later on, checking them on gel.
 +
<p class="experiment"></p>
 +
<p>Today a miniprep of pSB1K3 + J23100 + B0032 was performed. This gave concentrations of 51,3 and 89 ng/&mu;L.<br>
 +
<br>
 +
Of this, together with pSB1K3+ K352001 + B0015 and pSB1A3 a digest was performed:<br>
 +
<br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td>Biobricks</td>
 +
<td>Enzyme 1</td>
 +
<td>Enzyme 2</td>
 +
</tr>
 +
<tr>
 +
<td>1K3+J23+B32</td>
 +
<td>EcoRI</td>
 +
<td>SpeI</td>
 +
</tr>
 +
<tr>
 +
<td>1K3+K35+B15</td>
 +
<td>XbaI</td>
 +
<td>PstI</td>
 +
</tr>
 +
<tr>
 +
<td>pSB1A3</td>
 +
<td>EcoRI</td>
 +
<td>PstI</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<br>
 +
After two hours the digest was put on gel, though the rest was left for another two hours.<br>
 +
<br>
 +
&nbsp;
 +
<figure class="image"><img alt="" class="journalInlineImages" data-experimentfileid="2028112" data-img-journalid="1094404" id="img_file_id_2028112" src="/members/experiments/journal/authloader.ashx?fileID=2028112&amp;inline=true&amp;w=500">
 +
<figcaption>WhatsApp Image 2018-09-07 at 15.28.47.jpeg</figcaption>
 +
</figure>
 +
&nbsp;<br>
 +
&nbsp;
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</div>
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<div>
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<div class="notebook-image">
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<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/1/1f/T--Rotterdam_HR--Cooa_Experiment14.png');"></div>
 +
<p>CooA experiment 14</p>
 +
  </div>
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  </div>
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</div>
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 +
<div class="cardform">
 +
<div class="number"><b>14</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
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<h2 class="date" style="color: white;">September 6</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
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<h3 class="date">September 10</h3>
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<div class="attendees">Elise Grootscholten | Randall de Waard</div>
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<div class="description">
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<p>The minipreps of experiment 12 are digested with the restriction enzymes: SmaI and ScaI. Only Sca and Sma are both incubated at 37 degrees Celcius. Sma has to be incubated at 25 degrees Celcius. This is done in experiment 15.</p>
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 +
<p class="experiment"></p>
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<p><strong>Experiment 12: Digestions of the minipreps</strong><br>
 +
10-09-18<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
-SmaI (restriction-enzyme)<br>
 +
-ScaI (restriction-enzyme)<br>
 +
See materials experiment 10<br>
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<br>
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<strong>Methods</strong><br>
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See methods experiment 10<br>
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&nbsp;</p>
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<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
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<tbody>
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<tr>
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<td style="width: 221px;">DNA (A1 396.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
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<td style="width: 114px;">5.0&nbsp;&mu;L</td>
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</tr>
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<tr>
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<td style="width: 221px;">Cutsmart (10x)</td>
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<td style="width: 114px;">5&nbsp;&mu;L</td>
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</tr>
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<tr>
 +
<td style="width: 221px;">Sca1-HF (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
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<tr>
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<td style="width: 221px;">Sma1 (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
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<td style="width: 114px;">Fill to 50 &mu;L</td>
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</tr>
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</tbody>
 +
</table>
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<em>Table 1: Digestion scheme 1</em><br>
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&nbsp;
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<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
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<tbody>
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<tr>
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<td style="width: 221px;">DNA (B1 15.65 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
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<td style="width: 114px;">127.8</td>
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</tr>
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<tr>
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<td style="width: 221px;">Cutsmart (10x)</td>
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<td style="width: 114px;">5 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
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<td style="width: 114px;">Fill to 50 &mu;L</td>
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</tr>
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</tbody>
 +
</table>
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<em>Table 2: Digestion scheme 2</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (C1 186.6 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">10.7&nbsp;&mu;L</td>
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</tr>
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<tr>
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<td style="width: 221px;">Cutsmart (10x)</td>
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<td style="width: 114px;">5 &mu;L</td>
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</tr>
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<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
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<tr>
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<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
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</tr>
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</tbody>
 +
</table>
 +
&nbsp;<em>Table 3: Digestion scheme 3</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (D1 79.0&nbsp;&nbsp;&nbsp;&nbsp;ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">25.3 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
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<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
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<td style="width: 114px;">1 &mu;L</td>
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</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
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</tr>
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</tbody>
 +
</table>
 +
&nbsp;<em>Table 4: Digestion scheme 4</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (E1 97.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">20.7 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 5: Digestion scheme 5</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (F1 106.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">18.7 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 6: Digestion scheme 6</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (G1 328.9 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">6.1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 7: Digestion scheme 7</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (H1 121.0 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">16.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 8: Digestion scheme 8</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (I1 1129.5 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">&nbsp;1.7 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 9: Digestion scheme 9</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (J1 1465.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">&nbsp;1.4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 10: Digestion scheme 10</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (A2 310.4 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">6.4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 11: Digestion scheme 11</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (B2 364.9l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">5.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 12: Digestion scheme 12</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (C2 228.3 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">8.8 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 13: Digestion scheme 13</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (D2 701.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">2.9 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 14: Digestion scheme 14</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (E2 119.8 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">16.7 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 15: Digestion scheme 15</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (F2 493.3 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">4.1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 16: Digestion scheme 16</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (G2 211.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">9.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 17: Digestion scheme 17</em>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (H2 482.2 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">4.2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 18: Digestion scheme 18</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (I2 1364.7 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">1.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
&nbsp;<em>Table 19: Digestion scheme 19</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (J2 1665.5 ng/&mu;l)&nbsp;~ 2 &mu;g1.2</td>
 +
<td style="width: 114px;">&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF 1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1&nbsp; (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 20: Digestion scheme 20</em><br>
 +
<br>
 +
<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
The results of the digestions can be found in figure 1 and 2 (Images experiment 13)<br>
 +
<br>
 +
Ladder, A1, B1, C1, D1, E1, F1, G1, H1, I1, (J1), Ladder, A2, B2, C2, D2, E2, F2, G2, Ladder<br>
 +
Ladder, H2, I2, J2, Ladder, K3, J1, ladder<br>
 +
Something went wrong with the first (J1), so look at the second one.<br>
 +
<br>
 +
<strong>Conclusion/discussion</strong><br>
 +
Restriction-enzyme Sma was incubated at 37 degrees Celcius (same as Sca). This must be 25 degrees Celcius.<br>
 +
Therefore, Sma was added again and was incubated at 25 degrees Celcius. See results experiment 16.
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/1/1a/T--Rotterdam_HR--Gel1_exp13.jpeg');"></div>
 +
<p>Figure 1: Digestions with ScaI and SmaI</p>
 +
  </div>
 +
  <div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/5/5f/T--Rotterdam_HR--Gel2_exp13.jpeg');"></div>
 +
<p>Figure 2: Digestions with ScaI and SmaI</p>
 +
  </div>
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>13</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<svg viewBox="0 0 24 24">
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<h2 class="date" style="color: white;">September 10</h2>
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<h1 class="title">Soldering PCB [TC]</h1>
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<h3 class="date">September 11</h3>
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<label class="back" for="switch-102">&times;</label>
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</div>
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<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
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<div class="description">
 +
Testing first temperaturecontroller. Reworking surton components due to shorts on the PCB. Rework succesfull.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>03</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Soldering PCB [TC]</h3>
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<h1 class="title">Soldering PCB [TC]</h1>
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<h3 class="date">September 11</h3>
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<label class="back" for="switch-103">&times;</label>
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</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Testing second temperaturecontroller. Reworking surton components due to shorts on the PCB. Rework failed.
 +
<p class="experiment"></p>
 +
 +
</div>
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</div>
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<div class="cardform">
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<div class="number"><b>04</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Soldering PCB [TC]</h3>
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<h2 class="date" style="color: white;">September 11</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 13</h3>
 +
<label class="back" for="switch-104">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Minipreps of the biobrick combinations:&nbsp;K352002+K133116+pSB1K3 (1) and&nbsp;K352003+K133116+pSB1K3 (2) are done. The DNA concentration is for 1:189,3 ng/&mu;l and for 2: 355,3&nbsp;ng/&mu;l.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 14: Minipreps</strong></span><br>
 +
13-09-18<br>
 +
<br>
 +
Minipreps of the&nbsp;biobrick combinations:&nbsp;K352002+K133116+pSB1K3&nbsp;and&nbsp;K352003+K133116+pSB1K3<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
- Mini prep, Plasmid Purification Kit, Machery Nagel<br>
 +
- Eppendorf cups<br>
 +
- Variable Volume Pipets<br>
 +
- sterile pipet points<br>
 +
-Chloramphenicol (40 mg/mL)<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Machery Nagel Plasmid Purification<br>
 +
See experiment 3<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
K352002+K133116+pSB1K3: 189,3&nbsp;ng/&mu;l<br>
 +
K352003+K133116+pSB1K3: 355,3&nbsp;ng/&mu;l</p>
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 +
</div>
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<div>
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<div class="notebook-image">
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<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/parts/f/fa/T--Rotterdam_HR--Nanodrop_exp14.jpeg');"></div>
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<p>Figure 1: Minipreps 13-09-18</p>
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  </div>  
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<div class="cardform">
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<div class="number"><b>14</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<h2 class="date" style="color: white;">September 13</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 13</h3>
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<label class="back" for="switch-105">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Digestions have been done of the biobrickcombinations&nbsp;K352002 +K133116 + pSB1K3 and K352003 + K133116 + pSB1K3 with restriction-enzymes ScaI and SmaI</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Experiment 14: Digestions</strong><br>
 +
13-09-18<br>
 +
<br>
 +
Digestions of the biobrick combinations:&nbsp;K352002+K133116+pSB1K3&nbsp;and&nbsp;K352003+K133116+pSB1K3<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
-SmaI (restriction-enzyme)<br>
 +
-ScaI (restricion-enzyme)<br>
 +
See experiment..<br>
 +
<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
1&nbsp;&mu;L SmaI restriction-enzyme has been added to the digetions from experiment 13, except for pSB1K3.<br>
 +
After an our at room temperature the digestions were put in -20 degrees Celcius.</p>
 +
&nbsp;
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (K352002+K133116+pSB1K3 189,3 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">32,4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
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</tbody>
 +
</table>
 +
 +
<p><em>Table 1: Digestion scheme 1</em></p>
 +
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 346px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 221px;">DNA (K352003+K133116_pSB1K3 355,3 ng/&mu;l)&nbsp;~ 2 &mu;g</td>
 +
<td style="width: 114px;">5,6 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Cutsmart (10x)</td>
 +
<td style="width: 114px;">5&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sca1-HF (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Sma1 (2 U/&mu;L)</td>
 +
<td style="width: 114px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 221px;">Nuclease free water</td>
 +
<td style="width: 114px;">Fill to 50 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><em>Table 2: Digestion scheme 2</em></p>
 +
&nbsp;
 +
 +
<p>&nbsp;</p>
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</div>
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<div class="cardform">
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<div class="number"><b>15</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">September 13</h2>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 14</h3>
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<label class="back" for="switch-106">&times;</label>
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</div>
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<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>All the digestions of experiment 13 have digested again with SmaI for an our at 25 degrees Celcius. Because earlier the digestions have been put immediately bt 37 degrees Celcius. It looks like H1, J1 and D2 include the right biobricks.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 16: Gelelectroforesis</strong></span><br>
 +
14-09-18<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
-Loading dye (6x)<br>
 +
-2 Log DNA ladder<br>
 +
-Digested DNA<br>
 +
-1% arabinose gel (1 g/100 mL TAE buffer)<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Put 1 gram of arabinose with 100 mL TAE buffer.<br>
 +
Heat it in the microwave until the solution becomes clear.<br>
 +
Add 100 &mu;L gelred when the solution is hand warm.<br>
 +
Pour the solution into the gel holder which you taped before.<br>
 +
Wait until the gel is solid then you can pipette your dna with Loading dye into the wells.<br>
 +
<br>
 +
L: 3 &mu;L<br>
 +
DNA: 2&nbsp;&mu;L loading dye + 5 &mu;L DNA, 5 &mu;L on gel.<br>
 +
<br>
 +
<strong>Gel 1</strong><br>
 +
L, A1, B1, C1, D1, E1, F1, G1, G1 with only smaI, H1, I1, J1, A2, B2, C2, D2, E2, F2, G2, L<br>
 +
<br>
 +
<strong>Gel 2</strong><br>
 +
L, H2, I2, J2, L, K352002 + K133116 + pSB1K3,&nbsp; K352003&nbsp;+ K133116 + pSB1K3</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/parts/0/01/T--Rotterdam_HR--gel1exp16.jpeg');"></div>
 +
<p>Figure 1: Gel 1 (14-09-18), digested with ScaI and SmaI.</p>
 +
  </div>
 +
  <div class="notebook-image">
 +
<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/parts/d/d1/T--Rotterdam_HR--gel2exp16.jpeg');"></div>
 +
<p>Figure 2: Gel 2, H2, I2, J2, L, K352002+K133116+pSB1K3, K352003+K133116+pSB1K3</p>
 +
  </div>
 +
  </div>
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<div class="cardform">
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<div class="number"><b>16</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<svg viewBox="0 0 24 24">
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<h2 class="date" style="color: white;">September 14</h2>
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<h1 class="title">Testing gas production</h1>
 +
<h3 class="date">September 17</h3>
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</div>
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<div class="attendees">Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>Different&nbsp;strains&nbsp;of E.coli are tested as negative control. Those strains were:&nbsp;NEB10B&egrave;ta, BL21 (DE3), BL21, HB101, DH5alpha and&nbsp;JM109. NEB10B&egrave;ta produced the most gas. BL21, HB101 and JM101 produced none/almost none gas. Because BL21 isn't a K12 strain we can not use that one. That is why we will test further gasproduction in HB101 and JM109. And maybe those will be our final E.coli strains.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 5: Testing different strains&nbsp;of E.coli</strong></span><br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
All the following types of bacteria are E.coli:<br>
 +
-glycerolstock BL21<br>
 +
-glycerolstock HB101<br>
 +
-glycerolstock DH5alpha<br>
 +
-glycerolstock JM109<br>
 +
-agarplate with NEB10beta<br>
 +
-agarplat with BL21 (DE3)&nbsp;<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See experiment 1<br>
 +
Make overnight cultures of the following E.coli tribes: NEB10B&egrave;ta, BL21 (DE3), BL21, HB101, DH5alpha, JM109.<br>
 +
All the bacteria are de deluted in normal LB agar, LB agar with urea and LB agar with pyruvate.&nbsp;<br>
 +
<br>
 +
<br>
 +
<strong>Results</strong><br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td>Bacteria</td>
 +
<td>LB</td>
 +
<td>LB</td>
 +
<td>Urea</td>
 +
<td>Urea</td>
 +
<td>Pyruvate</td>
 +
<td>Pyruvate</td>
 +
</tr>
 +
<tr>
 +
<td>NEB10beta</td>
 +
<td>+</td>
 +
<td>+++</td>
 +
<td>+++</td>
 +
<td>+++</td>
 +
<td>+</td>
 +
<td>++/+++</td>
 +
</tr>
 +
<tr>
 +
<td>BL21 (DE3)&nbsp;</td>
 +
<td>-</td>
 +
<td>++</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
</tr>
 +
<tr>
 +
<td>BL21</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
</tr>
 +
<tr>
 +
<td>DH5a</td>
 +
<td>+++</td>
 +
<td>+/-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>+/-</td>
 +
<td>+</td>
 +
</tr>
 +
<tr>
 +
<td>HB101</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>+</td>
 +
<td>-</td>
 +
</tr>
 +
<tr>
 +
<td>JM109</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
<td>-</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing gas production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
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<label for="switch-107" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 17</h2>
 +
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">Designing peristaltic pump [PP]</h1>
 +
<h3 class="date">September 18</h3>
 +
<label class="back" for="switch-108">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Made a 3d-model of the parts for the peristaltic pump. Parts include: multiple gears, bearings, and casing.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing peristaltic pump [PP]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 18</h2>
 +
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 +
</svg>
 +
<h1 class="title">3d-printing peristaltic pump [PP]</h1>
 +
<h3 class="date">September 18</h3>
 +
<label class="back" for="switch-109">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
3d-printed all parts of the peristaltic pump model.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing peristaltic pump [PP]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-109" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 18</h2>
 +
</div>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Animating peristaltic pump [PP]</h1>
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<h3 class="date">September 18</h3>
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<label class="back" for="switch-110">&times;</label>
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</div>
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<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
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<div class="description">
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An animation was made of the first peristaltic pump.
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>03</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Animating peristaltic pump [PP]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-110" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 18</h2>
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</div>
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</div><style>
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}
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h1 class="title">Assessing CooA Production</h1>
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<h3 class="date">September 20</h3>
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<label class="back" for="switch-111">&times;</label>
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</div>
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<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
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<p>This week we peformed another cycle of cloning, starting with digesting J23105+B0032 with EcoRI and SpeI and K352001+B0015 with XbaI and PstI, checking these on gel (which seemed to be correct) and ligating them together on the pSB1A3 version of BBa_J04450. This was then grown overnight and prepped the following day.</p>
 +
 +
<p class="experiment"></p>
 +
<p>Today a dephos and ligation has been performd using the rAPId dephos and ligation kit and protocol.<br>
 +
For the dehpos a total of 17 &mu;L of pSB1A3 wasused, for an end concentration&nbsp;of 17 ng/&mu;L<br>
 +
For the ligationthe endvolume was trippled as to have enough DNA.<br>
 +
For both inserts (K352001 &amp; B0015 | J23100 + B0032)&nbsp;7.5 &mu;L was used (150 ng of DNA foreach insert)<br>
 +
For the veccor (pSB1A3) 3 &mu;L was used (51 ng of DNA)<br>
 +
The ligation will be left overnigh (started at 15:30).<br>
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<br>
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<br>
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&nbsp;</p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>15</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">September 20</h2>
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</div>
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#switch-112:checked ~ #card-112 .entryform {
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</style>
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<div class="card wetlab-card" id="card-112" onclick="">
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 24</h3>
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<label class="back" for="switch-112">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
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<div class="description">
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32 minipreps have been performed. A lot of them have a low concentration nucleic acid and/or have a high 260/280 and 260/230 rate. Therefore those will be done again.
 +
<p class="experiment"></p>
 +
<span style="font-size:14px;"><strong>Experiment 17: Minipreps</strong></span><br>
 +
20-09-18<br>
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Overnight cultures (35)<br>
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21-09-18<br>
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Minipreps (32)<br>
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<br>
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<strong>Materials&nbsp;</strong><br>
 +
See experiment 3<br>
 +
-Kanamycin (40 mg/mL)<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
See experiment 3<br>
 +
Make 32 minipreps:<br>
 +
A: 6<br>
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B: 6<br>
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C:6<br>
 +
D:1<br>
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E:6<br>
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F:6<br>
 +
H:1<br>
 +
I: 1<br>
 +
J:2<br>
 +
<br>
 +
<strong>Results/conclusion</strong><br>
 +
There was no grow in B6, C2 and C5.<br>
 +
A lot of them have a low concentration nucleic acid and/or have a high 260/280 and 260/230 rate. Therefore those will be done again.<br>
 +
Thise are: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5
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</div>
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<div>
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<div class="notebook-image">
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<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/parts/b/b9/T--Rotterdam_HR--nanodropexp17.jpeg');"></div>
 +
<p>Figure 1: Minipreps, a total of (32) because number 5, 7 and 12 are wrong.</p>
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  </div>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>17</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">September 24</h2>
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h1 class="title">Integration of silicon hose and peristaltic pump [PP]</h1>
 +
<h3 class="date">September 25</h3>
 +
<label class="back" for="switch-113">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Due to ordering issues a different silicon hose was implemented in the design. The new silicon tube was integrated into the 3d-printed peristaltic pump. A test was done with the new silicon tube in place. The test cocluded that the tube was not compatible with this peristaltic pump design. A new design must be made.
 +
<p class="experiment"></p>
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</div>
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</div>
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 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Integration of silicon hose and peristaltic pump [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 25</h2>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
 +
<h1 class="title">Designing peristaltic pump v2 [PP]</h1>
 +
<h3 class="date">September 25</h3>
 +
<label class="back" for="switch-114">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
Peristaltic pump was redesigned and the model has been updated.
 +
<p class="experiment"></p>
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 +
</div>
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</div>
 +
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing peristaltic pump v2 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-114" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 25</h2>
 +
</div>
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 +
</div><style>
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#switch-115:checked ~ #card-115 {
 +
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height: auto;
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 +
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 +
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 +
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display: none;
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</style>
 +
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<input type="checkbox" id="switch-115" class="switch"></input>
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<div class="entryform">
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<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">3d-printing peristaltic pump v2 [PP]</h1>
 +
<h3 class="date">September 25</h3>
 +
<label class="back" for="switch-115">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
3d-printed all parts of the peristaltic pump model v2. All parts have been put together succefully.
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing peristaltic pump v2 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-115" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 25</h2>
 +
</div>
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</div><style>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Testing peristaltic pump v2 [PP]</h1>
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<h3 class="date">September 25</h3>
 +
<label class="back" for="switch-116">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The silicon tube was intergrated with the 3e-printed peristaltic pump v2 and tested. The test concluded that the motor had insufficient force. A new design must be made.
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>04</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing peristaltic pump v2 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-116" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 25</h2>
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</div>
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</div><style>
 +
#switch-117:checked ~ #card-117 {
 +
width: 100%;
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height: auto;
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}
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#switch-117:checked ~ #card-117 .entryform {
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display: block;
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}
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#switch-117:checked ~ #card-117 .cardform,
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#switch-117:checked ~ #card-117::before,
 +
#switch-117:checked ~ #card-117::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-117" class="switch"></input>
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<div class="card hardware-card" id="card-117" onclick="">
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 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
 +
<h1 class="title">Designing peristaltic pump v3 [PP]</h1>
 +
<h3 class="date">September 26</h3>
 +
<label class="back" for="switch-117">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
An editional planetary gearset was added to the first planetary gearset of the peristaltic pump v2 model to increase the power output by 4x.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>01</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing peristaltic pump v3 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-117" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 26</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-118:checked ~ #card-118 {
 +
width: 100%;
 +
height: auto;
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}
 +
#switch-118:checked ~ #card-118 .entryform {
 +
display: block;
 +
}
 +
#switch-118:checked ~ #card-118 .cardform,
 +
#switch-118:checked ~ #card-118::before,
 +
#switch-118:checked ~ #card-118::after {
 +
display: none;
 +
}
 +
</style>
 +
 +
<input type="checkbox" id="switch-118" class="switch"></input>
 +
<div class="card hardware-card" id="card-118" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">3d-printing peristaltic pump v3 [PP]</h1>
 +
<h3 class="date">September 26</h3>
 +
<label class="back" for="switch-118">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
3d-printed all parts of the peristaltic pump model v3. All parts have been put together succefully.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing peristaltic pump v3 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 26</h2>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Testing peristaltic pump v3 [PP]</h1>
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<h3 class="date">September 26</h3>
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<label class="back" for="switch-119">&times;</label>
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</div>
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<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
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<div class="description">
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The newly pot together peristalticpump model v3 was tested. The test concluded a flaw in the design of the peristaltic pump v3. Failure: the silicon tube was pulled through the pump. A new design must be made.
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>03</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing peristaltic pump v3 [PP]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 26</h2>
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</div>
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</div><style>
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</style>
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<input type="checkbox" id="switch-120" class="switch"></input>
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<div class="card wetlab-card" id="card-120" onclick="">
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 +
<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h1 class="title">Assessing CooA Production</h1>
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<h3 class="date">September 26</h3>
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<label class="back" for="switch-120">&times;</label>
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</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
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Digesting the CooA combinations with NotI we could check if the requiered biobricks where present in our cells. Out of 10 checked colonies, 3 appeared to be the right size. Because of an error the on plate location of these colonies was lost however, so another check will have to be performed.
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<p class="experiment"></p>
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<p>Pipet 8 digestions<br>
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Nr. 1, 2, 4, 5, 7, 8, 9, and 10 of CooA. Use 5 &mu;L of digest with 3 &mu;L of loading dye.<br>
 +
<br>
 +
Gel:<br>
 +
Ladder 5 ul 1, 2, 4, 5, 7, 8, 9, 10, pSB1K3 (Sca), G1 (Sma), Ladder 5 ul, ladder 3 ul<br>
 +
The CooA digestions are&nbsp;5 ul<br>
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&nbsp;</p>
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 +
<figure class="image"><img alt="" class="journalInlineImages" data-experimentfileid="2101903" data-img-journalid="1128403" id="img_file_id_2101903" src="/members/experiments/journal/authloader.ashx?fileID=2101903&amp;inline=true&amp;w=500">
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<figcaption>Gel CooA.jpg</figcaption>
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</figure>
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<p>&nbsp;<br>
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&nbsp;</p>
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</div>
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<div>
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<div class="notebook-image">
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<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/3/38/T--Rotterdam_HR--CooA_18_09.jpeg');"></div>
 +
<p>CooA 18-09</p>
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  </div>  
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>16</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">September 26</h2>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Designing Test tube shaker [TTS]</h1>
 +
<h3 class="date">September 27</h3>
 +
<label class="back" for="switch-121">&times;</label>
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</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
A test tube shaker was designed to fit on top of a NEMU17 stepper motor. A spot for end switch was put in place to calibrate the test tube shaker. A cap for the test tube was designed.
 +
<p class="experiment"></p>
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</div>
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</div>
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 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing Test tube shaker [TTS]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 27</h2>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">3d-printing Test tube shaker [TTS]</h1>
 +
<h3 class="date">September 27</h3>
 +
<label class="back" for="switch-122">&times;</label>
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</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The test tube shaker was 3d-printed and assembled. Nine caps were printed.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing Test tube shaker [TTS]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 27</h2>
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
 +
<h1 class="title">Testing Test tube shaker [TTS]</h1>
 +
<h3 class="date">September 27</h3>
 +
<label class="back" for="switch-123">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The newly assembled testtube shaker was tested for sufficient and controlled shakage. The test was deemed a succes. The calibration for the test tube shaker potition was tested and deemed in working order. The caps were tested for sufficient air permeability. The test concluded suficient air permeability.
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>03</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing Test tube shaker [TTS]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 27</h2>
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<input type="checkbox" id="switch-124" class="switch"></input>
 +
<div class="card wetlab-card" id="card-124" onclick="">
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 27</h3>
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<label class="back" for="switch-124">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Making overnight cultures of the numbers: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5 with Kanamycin. These biobrick combinations can be found in experiment 12. Also&nbsp;the first digestions with XbaI, Eco0109I, HindIII and SspI-HF have been done.</p>
 +
 +
<p class="experiment"></p>
 +
<span style="font-size:14px;"><strong>Experiment 19: Making overnight cultures + first digestions</strong></span><br>
 +
27-09-18<br>
 +
<br>
 +
<strong>Materials</strong><br>
 +
-Kanamycine&nbsp;<br>
 +
-XbaI<br>
 +
-SspI-HF<br>
 +
-Eco019I<br>
 +
-HindIII<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Make new overnight cultures of the numbers:&nbsp;A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5 with Kanamycin.&nbsp;<br>
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<br>
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&nbsp;
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>19</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<label for="switch-124" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 27</h2>
 +
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">September 27</h3>
 +
<label class="back" for="switch-125">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Minipreps have been made of the numbers:&nbsp;A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5.</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 20: Minipreps of gasproduction biobricks</strong></span><br>
 +
27-09-18<br>
 +
<br>
 +
Make minipreps of A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5.<br>
 +
See experiment 3.<br>
 +
<br>
 +
Also make master mixes for the control digestions and code&nbsp;the tubes.<br>
 +
&nbsp;</p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>20</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
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<label for="switch-125" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 27</h2>
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<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
 +
<h1 class="title">Designing peristaltic pump v4 [PP]</h1>
 +
<h3 class="date">September 28</h3>
 +
<label class="back" for="switch-126">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The innershape of the tube duct was rounded to increase friction on the tube. The model of the peristaltic pump v3 was updated.
 +
<p class="experiment"></p>
 +
 +
</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing peristaltic pump v4 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 28</h2>
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</svg>
 +
<h1 class="title">3d-printing peristaltic pump v4 [PP]</h1>
 +
<h3 class="date">September 28</h3>
 +
<label class="back" for="switch-127">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The new parts for the tubeduct were 3d-printed and put together with the peritstaltic pump.
 +
<p class="experiment"></p>
 +
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing peristaltic pump v4 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-127" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 28</h2>
 +
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Testing peristaltic pump v4 [PP]</h1>
 +
<h3 class="date">September 28</h3>
 +
<label class="back" for="switch-128">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
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The new peristaltic pump v4 was tested. The test concluded a large improvement over the previous generation, but was deemed unreliable due tothe tube occasional being pulled through the pump. A new design must be made.
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>03</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing peristaltic pump v4 [PP]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-128" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 28</h2>
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</div>
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</div><style>
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display: none;
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</style>
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<input type="checkbox" id="switch-129" class="switch"></input>
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<div class="card hardware-card" id="card-129" onclick="">
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 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Designing peristaltic pump v4.1 [PP]</h1>
 +
<h3 class="date">September 28</h3>
 +
<label class="back" for="switch-129">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
A clamp was designed to hold the tube into the peristaltic pump from both ends. the peristaltic pump model v4 was updated.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing peristaltic pump v4.1 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-129" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 28</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-130:checked ~ #card-130 {
 +
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display: none;
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</style>
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<input type="checkbox" id="switch-130" class="switch"></input>
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<div class="card hardware-card" id="card-130" onclick="">
 +
 +
<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">3d-printing peristaltic pump v4.1 [PP]</h1>
 +
<h3 class="date">September 28</h3>
 +
<label class="back" for="switch-130">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The new clamp was 3d-printed, put together and added to the epristaltic pump.
 +
<p class="experiment"></p>
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</div>
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</div>
 +
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<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing peristaltic pump v4.1 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-130" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">September 28</h2>
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Testing peristaltic pump v4.1 [PP]</h1>
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<h3 class="date">September 28</h3>
 +
<label class="back" for="switch-131">&times;</label>
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</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The new peristaltic pump v4.1 was tested. The test conclude no noticable difference to the behaviour of the tube. The tube was still occsionally pulled through the pump. A new design must be made.
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<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>06</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing peristaltic pump v4.1 [PP]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-131" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 28</h2>
 +
</div>
 +
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</div><style>
 +
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 +
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</style>
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<input type="checkbox" id="switch-132" class="switch"></input>
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<div class="card hardware-card" id="card-132" onclick="">
 +
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<div class="entryform">
 +
<div class="titlebar">
 +
<svg viewBox="0 0 24 24">
 +
<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
 +
<h1 class="title">Designing peristaltic pump v5 [PP]</h1>
 +
<h3 class="date">September 30</h3>
 +
<label class="back" for="switch-132">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The tube clamp was attached to the top piece of the model's casing. The model of the persitaltic pump v4.1 was updated.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Designing peristaltic pump v5 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<label for="switch-132" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 30</h2>
 +
</div>
 +
 +
</div><style>
 +
#switch-133:checked ~ #card-133 {
 +
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 +
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<input type="checkbox" id="switch-133" class="switch"></input>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
 +
</svg>
 +
<h1 class="title">3d-printing peristaltic pump v5 [PP]</h1>
 +
<h3 class="date">September 30</h3>
 +
<label class="back" for="switch-133">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The new integrated tube clamp was 3d-printed and added to the peristaltic pump.
 +
<p class="experiment"></p>
 +
 +
</div>
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</div>
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<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">3d-printing peristaltic pump v5 [PP]</h3>
 +
<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<label for="switch-133" class="more-info">Show more</label>
 +
<h2 class="date" style="color: white;">September 30</h2>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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</svg>
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<h1 class="title">Testing peristaltic pump v5 [PP]</h1>
 +
<h3 class="date">September 30</h3>
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<label class="back" for="switch-134">&times;</label>
 +
</div>
 +
<div class="attendees">Tom van Dijk | Mike Vrieswijk</div>
 +
<div class="description">
 +
The new peristaltic pump v5 was tested.  The new v5 was deemed a succes. No previous bad behaviour was observed.
 +
<p class="experiment"></p>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>03</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing peristaltic pump v5 [PP]</h3>
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<svg viewBox="0 0 24 24">
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<path d="M15.9,18.45C17.25,18.45 18.35,17.35 18.35,16C18.35,14.65 17.25,13.55 15.9,13.55C14.54,13.55 13.45,14.65 13.45,16C13.45,17.35 14.54,18.45 15.9,18.45M21.1,16.68L22.58,17.84C22.71,17.95 22.75,18.13 22.66,18.29L21.26,20.71C21.17,20.86 21,20.92 20.83,20.86L19.09,20.16C18.73,20.44 18.33,20.67 17.91,20.85L17.64,22.7C17.62,22.87 17.47,23 17.3,23H14.5C14.32,23 14.18,22.87 14.15,22.7L13.89,20.85C13.46,20.67 13.07,20.44 12.71,20.16L10.96,20.86C10.81,20.92 10.62,20.86 10.54,20.71L9.14,18.29C9.05,18.13 9.09,17.95 9.22,17.84L10.7,16.68L10.65,16L10.7,15.31L9.22,14.16C9.09,14.05 9.05,13.86 9.14,13.71L10.54,11.29C10.62,11.13 10.81,11.07 10.96,11.13L12.71,11.84C13.07,11.56 13.46,11.32 13.89,11.15L14.15,9.29C14.18,9.13 14.32,9 14.5,9H17.3C17.47,9 17.62,9.13 17.64,9.29L17.91,11.15C18.33,11.32 18.73,11.56 19.09,11.84L20.83,11.13C21,11.07 21.17,11.13 21.26,11.29L22.66,13.71C22.75,13.86 22.71,14.05 22.58,14.16L21.1,15.31L21.15,16L21.1,16.68M6.69,8.07C7.56,8.07 8.26,7.37 8.26,6.5C8.26,5.63 7.56,4.92 6.69,4.92A1.58,1.58 0 0,0 5.11,6.5C5.11,7.37 5.82,8.07 6.69,8.07M10.03,6.94L11,7.68C11.07,7.75 11.09,7.87 11.03,7.97L10.13,9.53C10.08,9.63 9.96,9.67 9.86,9.63L8.74,9.18L8,9.62L7.81,10.81C7.79,10.92 7.7,11 7.59,11H5.79C5.67,11 5.58,10.92 5.56,10.81L5.4,9.62L4.64,9.18L3.5,9.63C3.41,9.67 3.3,9.63 3.24,9.53L2.34,7.97C2.28,7.87 2.31,7.75 2.39,7.68L3.34,6.94L3.31,6.5L3.34,6.06L2.39,5.32C2.31,5.25 2.28,5.13 2.34,5.03L3.24,3.47C3.3,3.37 3.41,3.33 3.5,3.37L4.63,3.82L5.4,3.38L5.56,2.19C5.58,2.08 5.67,2 5.79,2H7.59C7.7,2 7.79,2.08 7.81,2.19L8,3.38L8.74,3.82L9.86,3.37C9.96,3.33 10.08,3.37 10.13,3.47L11.03,5.03C11.09,5.13 11.07,5.25 11,5.32L10.03,6.06L10.06,6.5L10.03,6.94Z"></path>
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<h2 class="date" style="color: white;">September 30</h2>
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</div>
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<div class="card wetlab-card" id="card-135" onclick="">
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<div class="entryform">
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">October 1</h3>
 +
<label class="back" for="switch-135">&times;</label>
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</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
Today a colony PCR was performed of multiple CooA ligation products of J23100+B0032+K352001+B0015 on pSB1A3. This specific colony was later PCRed again with the gasproduction biobricks in preperation for sequencing. Only one of the colonies seemed to be the right length, though te lenght of the complete insert and BBa_J04450 is quite similair, so sequencing later this week will provide the final answer.<br>
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<br>
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&nbsp;
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<p class="experiment"></p>
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</div>
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<div class="notebook-image">
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<div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/9/94/T--Rotterdam_HR--PCR_Producten_4_10.jpeg');"></div>
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<p>CooA 4-10</p>
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  </div>  
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</div>
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<div class="cardform">
 +
<div class="number"><b>17</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">October 1</h2>
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<div class="titlebar">
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
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<h3 class="date">October 4</h3>
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<label class="back" for="switch-136">&times;</label>
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</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>From 27-09 until 04-10 more than 200 digestions and 17 gelelectrophoresis have been performed. This was done as a control for our composite parts. We digested with XbaI, Eco01019 and SspI-HF/HindIII. Every restriction-enzyme has an unique place to cut in our constructs. With this we could test if the promoter, backbone and gas production gene was present. However, the digestions didn't show us very good results. Only I1 and J1 looked good.&nbsp;After this we started with&nbsp;PCR.</p>
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<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 21: Digestions and gelelectrophoresis</strong></span><br>
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01-10-18<br>
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<br>
 +
Digestions of the gasproduction biobrick combinations (D, H, I and J) and a start with the gelelectrophoresis.<br>
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<br>
 +
<strong>Methods</strong><br>
 +
Digestions of D2, H1, I2 and J2 of experiment 12.<br>
 +
J2 will from now on be J3.</p>
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<table border="1" cellpadding="1" cellspacing="1">
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<tbody>
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<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>4 &mu;L</td>
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</tr>
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<tr>
 +
<td>Eco0109I (20 U/&mu;L)</td>
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<td>1 &mu;L</td>
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</tr>
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<tr>
 +
<td>Nuclease free water</td>
 +
<td>35 &mu;L</td>
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</tr>
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</tbody>
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</table>
 +
 +
<p><em>Table 1: Master mix digestion 1</em><br>
 +
&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Eco0109I (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>34 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 2: Master mix digestion 2</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Eco0109I (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>HindIII&nbsp;(20 U/&mu;L)</td>
 +
<td>1&nbsp;&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>15 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 3: Master mix digestion 3</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Eco0109I (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>SspI-HF</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>15&nbsp; &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 4: Master mix digestion 4</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>HindIII (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>16 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 5: Master mix digestion 5</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>SspI-HF (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>16 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 6: Master mix digestion 6</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>XbaI (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>35 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 7: Master mix digestion 7</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>HindIII (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>17 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 8: Master mix digestion 8</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>SspI-HF (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>17 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 9: Master mix digestion 9</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Eco0109I (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>HindIII (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>16 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 10: Master mix digestion 10</em><br>
 +
&nbsp;
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>NEB 2.1 (10x)</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Eco0109I (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>SspI-HF (20 U/&mu;L)</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nuclease free water</td>
 +
<td>16 &mu;L<br>
 +
&nbsp;</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<em>Table 11: Master mix digestion 11</em><br>
 +
<br>
 +
Pipette 9&nbsp;&mu;L&nbsp;of the mastermix with 2&nbsp;&mu;L&nbsp;of H1/ 1&nbsp;&mu;L&nbsp;of D2/ 1 &mu;L&nbsp;of I2/ 1 &mu;L&nbsp;of J23<br>
 +
<br>
 +
D2 and J3 are digested with mastermix: 1, 2, 3, 5, 7, 8 and 10.<br>
 +
H1 and I2 are digested with mastermix: 1, 2, 4, 6, 7, 9, and 11.<br>
 +
<br>
 +
L= 2 log DNA ladder<br>
 +
&nbsp;
 +
<p><strong>Gelelectrophoresis</strong><br>
 +
01-10-18<br>
 +
<br>
 +
<strong>Gel 1</strong><br>
 +
L, B6 Eco01090I, B6 SspI-HF, B6 XbaI, B6 Eco0109I + SspI-HF, B6 Eco0109I + XbaI, B6 SspI-HF + XbaI, B6 Eco0109I + SspI-HF + XbaI, L, B2 SspI-HF, B2 XbaI, B2 Eco0109I + SspI-HF, B2 Eco0109I + XbaI, B2 SspI-HF + XbaI, B2 Eco0109I + SspI-HF + XbaI, L, H1 SspI-HF, H1 XbaI, H1Eco0109I + SspI-HF, H1 Eco0109I + XbaI<br>
 +
<br>
 +
<strong>Gel 2</strong><br>
 +
L, H1 SspI-HF + XbaI, H1 Eco0109I + SspI-HF + XbaI, I1 Eco0109I + SspI-HF, I1 Eco0109I + XbaI, I1 SspI-HF + XbaI, I1 Eco0109I + SspI-HF + XbaI, L, I1 SspI-HF, I1 XbaI, B3 SspI-HF, B3 XbaI, B3 Eco0109I + SspI-HF, B3 Eco0109I + XbaI, B3 SspI-HF + XbaI, B4 SspI-HF, B4 XbaI, B4 Eco0109I + SspI-HF, B4 Eco0109I + XbaI, B4 Eco0109I + SspI-HF + XbaI<br>
 +
<br>
 +
<strong>Gel 3</strong><br>
 +
L, F6 SspI-HF, F6 XbaI, F6 Eco0109I + SspI-HF, F6 Eco0109I + XbaI, F6 SspI-HF + XbaI, F6 Eco0109I + SspI-HF + XbaI, L, F5 SspI-HF, F5 XbaI, F5 Eco0109I + SspI-HF, F5 Eco0109I + XbaI, F5 SspI-HF + XbaI, F5 Eco0109I + SspI-HF + XbaI, L F4 XbaI, F4 Eco0109I + SspI-HF, F4 Eco0109I + XbaI, F4 sspI-HF + XbaI, L<br>
 +
<br>
 +
<strong>Gel 4</strong><br>
 +
L, F4 Eco0109 + XbaI + SspI-HF, F4 Eco0109, F3&nbsp;XbaI, F3&nbsp;Eco0109 + SspI-HF, F3 Eco0109 + XbaI, F3 SspI-HF + XbaI, F3 Eco0109 + XbaI + SspI-HF, F2 SspI-HF, F2 XbaI, L, F2 Eco0109 + SspI-HF, F2 Eco0109 + XbaI, F2 SspI-HF + XbaI, F2 Eco01090 + XbaI + SspI-HF, F1 XbaI, F1 Eco0109 + SspI-HF, F1 Eco0109 + XbaI, F1 SspI-HF + XbaI<br>
 +
<br>
 +
<strong>Gel 5</strong><br>
 +
L, F1 Eco0109 + XbaI + SspI-HF, E1 Eco0109, E1 SspI-HF, E1 XbaI, E1 Eco0109 + SspI-HF, E1 Eco0109 + XbaI, E1 SspI-HF + XbaI, E1 Eco0109 + XbaI + SspI-HF, L,&nbsp;E2&nbsp;Eco0109, E2&nbsp;SspI-HF, E1 XbaI, E2&nbsp;Eco0109 + SspI-HF, E2&nbsp;Eco0109 + XbaI, E2&nbsp;SspI-HF + XbaI, E2&nbsp;Eco0109 + XbaI + SspI-HF, L<br>
 +
<br>
 +
<strong>Gel 6</strong><br>
 +
L,&nbsp;E3&nbsp;SspI-HF, E3&nbsp;XbaI, E3&nbsp;Eco0109 + SspI-HF, E3&nbsp;Eco0109 + XbaI, E3&nbsp;SspI-HF + XbaI, E3&nbsp;Eco0109 + XbaI + SspI-HF, L,&nbsp;E6&nbsp;Eco0109, E6&nbsp;SspI-HF, E6&nbsp;XbaI, E6&nbsp;Eco0109 + SspI-HF, E6&nbsp;Eco0109 + XbaI, E6&nbsp;SspI-HF + XbaI, E6&nbsp;Eco0109 + XbaI + SspI-HF, L<br>
 +
<br>
 +
02-10-18<br>
 +
<strong>Gelelectrophoresis</strong><br>
 +
<br>
 +
<strong>Gel 1</strong><br>
 +
L,- , A1 Eco0109 , A1 SspI-HF, A1 XbaI, A1 Eco0109 + SspI-HF, A1 Eco01019 + XbaI, A1 SspI-HF + XbaI, A1 Eco0109 + XbaI + SspI-HF, -, -,&nbsp;A2&nbsp;Eco0109 , A2&nbsp;SspI-HF, A2&nbsp;XbaI, A2&nbsp;Eco0109 + SspI-HF, A2&nbsp;Eco01019 + XbaI, A2&nbsp;SspI-HF + XbaI, A2&nbsp;Eco0109 + XbaI + SspI-HF, -, L<br>
 +
<br>
 +
<strong>Gel 2</strong><br>
 +
L,- , A3&nbsp;Eco0109 , A3&nbsp;SspI-HF, A3&nbsp;XbaI, A3&nbsp;Eco0109 + SspI-HF, A3&nbsp;Eco01019 + XbaI, A3&nbsp;SspI-HF + XbaI, A3&nbsp;Eco0109 + XbaI + SspI-HF, -, -,&nbsp;A4&nbsp;Eco0109 , A4&nbsp;SspI-HF, A4&nbsp;XbaI, A4&nbsp;Eco0109 + SspI-HF, A4&nbsp;Eco01019 + XbaI, A4&nbsp;SspI-HF + XbaI, A4&nbsp;Eco0109 + XbaI + SspI-HF, -, L<br>
 +
<br>
 +
<br>
 +
03-10-18<br>
 +
<strong>Gelelectrophoresis</strong><br>
 +
<br>
 +
<strong>Gel 1</strong><br>
 +
L, D2 Eco0109, D2 Eco0109 + XbaI, D2 Eco0109 +XbaI + HindIII, D2 HindII + XbaI, D2 XbaI, D2 HindIII, D2 Eco0109 + HindIII, D not digested, L, J3 Eco0109, J3 Eco0109 + XbaI, J3 Eco0109 + XbaI + HindIII, J3 XbaI, J3 HindIII, J3 Eco0109 + HindIII, J3.10, J not digested, L.<br>
 +
<br>
 +
<strong>Gel 2</strong><br>
 +
L, H2 Eco0109, H2 Eco0109 + XbaI,&nbsp;H2 Eco0109 + XbaI + SspI-HF, H2 SspI-HF + XbaI, H2 XbaI, H2 SspI-HF, H2 Eco0109 + SspI-HF, H2 not digested, L, I2 Eco0109, I2 Eco0109 + XbaI, I2 Eco0109 + XbaI + SspI-HF, I2 SspI-HF + XbaI, I2 XbaI, I2 SspI-HF, I2 Eco0109 +SspI-HF, I2 not digested, L<br>
 +
<br>
 +
<strong>Gel 3</strong><br>
 +
L, C1 Eco0109, C1 HindIII, C1 XbaI, C1 Eco0109 + XbaI, C1 Eco0109 + HindIII, C1&nbsp;HindIII + XbaI, C1 Eco0109 + XbaI + HindIII, C1 not digested, L, C2 Eco0109, C2 HindIII, C2 XbaI, C2 Eco0109 + XbaI, C2 Eco0109 + HindIII, C2 HindIII + XbaI, C2 Eco0109 + XbaI + HindIII, C2 not dgested, L<br>
 +
<br>
 +
<strong>Gel 4</strong><br>
 +
L, C3&nbsp;Eco0109, C3&nbsp;HindIII, C3&nbsp;XbaI, C3&nbsp;Eco0109 + XbaI, C3&nbsp;Eco0109 + HindIII, C3&nbsp;HindIII + XbaI, C3&nbsp;Eco0109 + XbaI + HindIII, C3&nbsp;not digested, L, C4&nbsp;Eco0109, C4&nbsp;HindIII, C4&nbsp;XbaI, C4&nbsp;Eco0109 + XbaI, C4&nbsp;Eco0109 + HindIII, C4&nbsp;HindIII + XbaI, C4&nbsp;Eco0109 + XbaI + HindIII, C4&nbsp;not dgested, L<br>
 +
<br>
 +
<strong>Gel 5</strong><br>
 +
L, C6 Eco0109, C6 HindIII, C6 XbaI, C6 Eco0109 + XbaI, C6 Eco0109 + HindIII, C6 HindIII + XbaI, C6 Eco0109 + SspI-HF + XbaI, C6 not digested, L, B5 Eco0109, B5 SspI-HF, B5 XbaI, B5 XbaI&nbsp;(other tube), B5 Eco0109 + XbaI, B5 SspI-HF + Eco0109, B5 SspI-HF + XbaI, B5 Eco0109 + SspI-HF + XbaI, L<br>
 +
<br>
 +
04-10-18<br>
 +
<strong>Gelelectrophoresis</strong><br>
 +
<br>
 +
<strong>Gel 1</strong><br>
 +
L, J2 Eco0109, J2 HindIII, J2 XbaI, J2 Eco0109 + XbaI, J2 Eco0109 + HindIII, J2 HindIII + XbaI, J2 Eco0109 + HindIII + XbaI, J2 not digested, L,&nbsp;J1&nbsp;Eco0109, J1&nbsp;HindIII, J1&nbsp;XbaI, J1&nbsp;Eco0109 + XbaI, J1&nbsp;Eco0109 + HindIII, J1&nbsp;HindIII + XbaI, J1&nbsp;Eco0109 + HindIII + XbaI, J1&nbsp;not digested, L<br>
 +
<br>
 +
<strong>Gel 2</strong><br>
 +
L, B2 Eco0109, B2 HindIII, B3 Eco0109, B3 HindIII, B3 Eco0109 + SspI-HF + XbaI, B4 Eco0109, B4 HindIII, B4 Eco0109 + SspI-HF + XbaI, not digested B2, L, D1 Eco0109, D1 HindIII, D1 XbaI, D1 Eco0109 + XbaI, D1 Eco0109 + HindIII, D1 HindIII + XbaI, D1 Eco0109 + HindIII + XbaI, not digested D1, L<br>
 +
<br>
 +
<strong>Gel 3</strong><br>
 +
L, F1 Eco0109, F1 HindIII, F2 Eco0109, F3 Eco0109, F3 HindIII, F6 Eco0109, F6 HindIII not digested F6, L, A2 HindIII, A3 HindIII, not digested A1, E3 Eco0109, E3 HindIII, not digested E3, L<br>
 +
<br>
 +
<strong>Gel 4</strong><br>
 +
L, H1 Eco0109, H1 HindIII, H1 not digested, I1 Eco0109, I1 HindIII, L</p>
 +
 +
</div>
 +
<div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/fb/T--Rotterdam_HR--gel1_01_10.jpeg');"></div>
 +
  <p>Figure 1: Gel 1 01-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/4/43/T--Rotterdam_HR--Gel2_01_10.jpeg');"></div>
 +
  <p>Figure 2: Gel 2 01-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/c/c5/T--Rotterdam_HR--Gel301-10.jpeg');"></div>
 +
  <p>Figure 3: Gel 3 01-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/4/48/T--Rotterdam_HR--Gel4_01_10.jpeg');"></div>
 +
  <p>Figure 4: Gel 4 01-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/6/6f/T--Rotterdam_HR--Gel5_01_10.jpeg');"></div>
 +
  <p>iFigure 5: Gel 5 01-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/5/5a/T--Rotterdam_HR--Gel6_01_10.jpeg');"></div>
 +
  <p>iFigure 6: Gel 6 01-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/fd/T--Rotterdam_HR--gel2_10_A1A2.jpeg');"></div>
 +
  <p>Figure 7: Gel 1 02-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/2/26/T--Rotterdam_HR--gel02_10_A3A4.jpeg');"></div>
 +
  <p>Figure 8: Gel 2 02-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/c/ca/T--Rotterdam_HR--Gel1_03_10.jpeg');"></div>
 +
  <p>Figure 9: Gel 1 03-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/d/d6/T--Rotterdam_HR--Gel2_03_10.jpeg');"></div>
 +
  <p>Figure 10: Gel 2 03-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/0/0f/T--Rotterdam_HR--Gel3_03_10.jpeg');"></div>
 +
  <p>Figure 11: Gel 3 03-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/0/08/T--Rotterdam_HR--Gel4_03_10.jpeg');"></div>
 +
  <p>Figure 12: Gel 4 03-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/9/95/T--Rotterdam_HR--Gel5_03_10.jpeg');"></div>
 +
  <p>Figure 13: Gel 5 03-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/0/03/T--Rotterdam_HR--Gel1_04_10.jpeg');"></div>
 +
  <p>Figure 14: Gel 1 04-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/1/1c/T--Rotterdam_HR--Gel2_04_10.jpeg');"></div>
 +
  <p>Figure 15: Gel 2 04-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/c/cc/T--Rotterdam_HR--Gel3_04_10.jpeg');"></div>
 +
  <p>Figure 16: Gel 3 04-10-18</p>
 +
</div>
 +
<div class="notebook-image">
 +
  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/f/fa/T--Rotterdam_HR--Gel4_04_10.jpeg');"></div>
 +
  <p>Figure 17: Gel 4 04-10-18</p>
 +
</div>
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>21</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
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 +
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 +
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 +
<h2 class="date" style="color: white;">October 4</h2>
 +
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</svg>
 +
<h1 class="title">Preparing DNA for Submission</h1>
 +
<h3 class="date">October 5</h3>
 +
<label class="back" for="switch-137">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin Vermeulen | Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Today we made colony PCRs of the biobricks we suspected that where correct of our gasproduction, and of colony 8 of CooA. These where then send to the Erasmus Medical Centre, who sequenced our parts for us.</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Colony PCR</strong><br>
 +
Materials: For our PCR (Both plasmid and colony) reactions we have used the standard neb protocols for a 25&nbsp;&mu;l reaction , with the following concentrations:<br>
 +
<br>
 +
10X Standard Taq Reaction Buffer&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;2.5&nbsp;&mu;l<br>
 +
10 mM dNTPs&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;0.5&nbsp;&mu;l<br>
 +
10&nbsp;&mu;M forward Primer&nbsp; (VF2)&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;0.5&nbsp;&mu;l<br>
 +
10&nbsp;&mu;M reverse Primers&nbsp; (VR)&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;0.5&nbsp;&mu;l<br>
 +
Taq DNA Polymerase&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp; &nbsp;0.125 ul<br>
 +
Nuclease-free water&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;to 25&nbsp;&mu;l<br>
 +
<br>
 +
The primers VF2 and VR got delivered in powder form. So it was dissolved in Nuclease-free water till a concentration of 100&nbsp;&mu;m.<br>
 +
<br>
 +
Methods: A master mix is made with the concentrations shown above. In case of a plasmid pcr, a volume of plasmid is added before the reaction is filled with Nuclease-free water. With the colony PCR we used a toothpick to touch on the desired colony. Before briefly submerging and shaking the tip of the toothpick in the PCR Eppendorfcup filled with the mastermix. Before pricking it into a clean&nbsp;Agar plate a couple of times to grow more colonies in case the PCR shows us the desired products<br>
 +
<br>
 +
For our PCR reactions we use the Bio-Rad T100 thermal cycler. Afterwards the products of the PCR reaction were ran on a gel electrophoresis.<br>
 +
<br>
 +
<strong>PCR Purification</strong><br>
 +
8 &mu;L PCR product&nbsp;+ 2 &mu;L ExoSAP-IT<br>
 +
37&deg;C 20 minutes<br>
 +
80&deg;C 15 minutes<br>
 +
<br>
 +
&nbsp;</p>
 +
 +
<p><strong>Sequencing Reaction</strong></p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1">
 +
<tbody>
 +
<tr>
 +
<td>4,0 &mu;L</td>
 +
<td>PCR product</td>
 +
</tr>
 +
<tr>
 +
<td>0,15 &mu;L</td>
 +
<td>forward or reverse primer</td>
 +
</tr>
 +
<tr>
 +
<td>0,5 &mu;L</td>
 +
<td>BigDye V3.1 (Applied Biosystems)</td>
 +
</tr>
 +
<tr>
 +
<td>1,75 &mu;L</td>
 +
<td>Sequencing buffer (5x)</td>
 +
</tr>
 +
<tr>
 +
<td>3,6 &mu;L</td>
 +
<td>Nuclease free water</td>
 +
</tr>
 +
<tr>
 +
<td>10 &mu;L</td>
 +
<td>Total</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><br>
 +
96&deg;C&nbsp;1 minute<br>
 +
96&deg;C&nbsp;10 seconds<br>
 +
55&deg;C 5 seconds<br>
 +
60&deg;C 2 minutes<br>
 +
run for 35 cycli<br>
 +
10&deg;C&nbsp;end<br>
 +
<br>
 +
<strong>Sequencing reaction purification</strong><br>
 +
Following protocol of CleanSeq Agencourt<br>
 +
<br>
 +
<strong>Sequencing</strong><br>
 +
ABI 3730 XL, 50 cm capillair</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>01</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Preparing DNA for Submission</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">October 5</h2>
 +
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Preparing DNA for Submission</h1>
 +
<h3 class="date">October 7</h3>
 +
<label class="back" for="switch-138">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin Vermeulen | Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Today we digested our succesfully sequenced biobricks (B1 / B6 (same biobrick), D1, I1 and J2), using EcoRI and PstI to get them into a pSB1C3 backbone. This was then transormed and plated. And the end of the day we made 6 overnight cultures of the barely visible colonies.</p>
 +
 +
<p class="experiment"></p>
 +
<p><strong>Materials</strong><br>
 +
-Restriction enzymes EcoRI-HF and PstI<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
Digestion<br>
 +
<br>
 +
Mastermix</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 184px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 114px;">Cutsmart (10x)</td>
 +
<td style="width: 57px;">55 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 114px;">EcoRI-HF (20 U/&mu;L)</td>
 +
<td style="width: 57px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 114px;">PstI (20 U/&mu;L)</td>
 +
<td style="width: 57px;">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 114px;">Nuclease free water</td>
 +
<td style="width: 57px;">382,7 &mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><br>
 +
50&nbsp;&mu;L mastermix&nbsp;- ...&nbsp;&mu;L DNA (see below)<br>
 +
A2: 12,6&nbsp;&mu;L<br>
 +
B1: 15,7&nbsp;&mu;L<br>
 +
B5: 9,4&nbsp;&mu;L<br>
 +
B6: 9,6&nbsp;&mu;L<br>
 +
D1: 10,2&nbsp;&mu;L<br>
 +
E1: 15,7&nbsp;&mu;L (1&nbsp;&mu;g)<br>
 +
I1: 14,3&nbsp;&mu;L (1 &mu;g)<br>
 +
J1.2: 17,4&nbsp;&mu;L<br>
 +
CooA (8): 5,4&nbsp;&mu;L<br>
 +
<br>
 +
Dephosphorylation + ligation using rAPId dephos and ligation protocol and kit</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Preparing DNA for Submission</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h2 class="date" style="color: white;">October 7</h2>
 +
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Preparing DNA for Submission</h1>
 +
<h3 class="date">October 9</h3>
 +
<label class="back" for="switch-139">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin Vermeulen | Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Today we mini prepped our DNA (~6 colonies per succesfully sequenced biobrick, overnight cultures made on the 8th of october) and digested this.</p>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>03</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Preparing DNA for Submission</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h2 class="date" style="color: white;">October 9</h2>
 +
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
<h1 class="title">Testing gas production</h1>
 +
<h3 class="date">October 11</h3>
 +
<label class="back" for="switch-140">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>Overnight cultures made of the following strains with the right plasmid:</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 322px;">
 +
<tbody>
 +
<tr>
 +
<td style="width: 88px;"><strong>Number</strong></td>
 +
<td style="width: 88px;"><strong>BBa_</strong></td>
 +
<td style="width: 105px;"><strong>HB101</strong></td>
 +
<td style="width: 113px;"><strong>JM109</strong></td>
 +
</tr>
 +
<tr>
 +
<td style="width: 88px;">J1</td>
 +
<td style="width: 88px;">BBa_K2604000</td>
 +
<td style="width: 105px;">1x</td>
 +
<td style="width: 113px;">1x</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 88px;">I1</td>
 +
<td style="width: 88px;">BBa_K2604001</td>
 +
<td style="width: 105px;">3x</td>
 +
<td style="width: 113px;">3x</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 88px;">D1</td>
 +
<td style="width: 88px;">BBa_K2604003</td>
 +
<td style="width: 105px;">2x</td>
 +
<td style="width: 113px;">2x</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 88px;">B1</td>
 +
<td style="width: 88px;">BBa_K2604006</td>
 +
<td style="width: 105px;">1x</td>
 +
<td style="width: 113px;">1x</td>
 +
</tr>
 +
<tr>
 +
<td style="width: 88px;">B6</td>
 +
<td style="width: 88px;">BBa_K2604006</td>
 +
<td style="width: 105px;">1x</td>
 +
<td style="width: 113px;">1x</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p><em>Table 1: Overnight culture scheme.</em><br>
 +
<br>
 +
I1, D1 and J1 are tested for the gas productio. The rest is only made for glycerolstocks<br>
 +
<br>
 +
We can not say for sure if the PBad promoter and the gas production enzyme pyruvate decarboxylase work. Fot that more gas production test have to be done. Also the negative controls produce gas. This is a problem. The JM109 and HB101 E.coli strains didn't produce gas in experiment .&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 6: Testing gasproduction with our new biobricks.</strong></span><br>
 +
<br>
 +
Testing our new biobricks K260401 in JM109 and HB101. As a negative control we used the other new biobricks ...<br>
 +
<br>
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<strong>Materials</strong><br>
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-LB culture medium<br>
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-Kanamycin<br>
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-Bacteria on agar plates with the right plasmid<br>
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<br>
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<strong>Methods</strong><br>
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Make overnight cultures of the following strains with the right plasmid:<br>
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&nbsp;</p>
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<table border="1" cellpadding="1" cellspacing="1" style="width: 322px;">
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<tbody>
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<tr>
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<td style="width: 88px;"><strong>Number</strong></td>
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<td style="width: 88px;"><strong>BBa_</strong></td>
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<td style="width: 105px;"><strong>HB101</strong></td>
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<td style="width: 113px;"><strong>JM109</strong></td>
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</tr>
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<tr>
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<td style="width: 88px;">J1</td>
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<td style="width: 88px;">BBa_K2604000</td>
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<td style="width: 105px;">1x</td>
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<td style="width: 113px;">1x</td>
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</tr>
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<tr>
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<td style="width: 88px;">I1</td>
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<td style="width: 88px;">BBa_K2604001</td>
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<td style="width: 105px;">3x</td>
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<td style="width: 113px;">3x</td>
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</tr>
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<tr>
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<td style="width: 88px;">D1</td>
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<td style="width: 88px;">BBa_K2604003</td>
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<td style="width: 105px;">2x</td>
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<td style="width: 113px;">2x</td>
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</tr>
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<tr>
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<td style="width: 88px;">B1</td>
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<td style="width: 88px;">BBa_K2604006</td>
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<td style="width: 105px;">1x</td>
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<td style="width: 113px;">1x</td>
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</tr>
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<tr>
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<td style="width: 88px;">B6</td>
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<td style="width: 88px;">BBa_K2604006</td>
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<td style="width: 105px;">1x</td>
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<td style="width: 113px;">1x</td>
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</tr>
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</tbody>
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</table>
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<em>Table 1: Overnight culture scheme.</em><br>
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<br>
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Only the numbers J1, I1 and D1 are tested. The rest is made for making glycerolstocks.<br>
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J1 and D1 are the negative controls in this experiment.<br>
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<br>
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LB: 582 mL +18 mL 100 mM sodiumpyruvate and 525 &mu;L kanamycin (40 mg/mL).<br>
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After this the LB is devided in 2. Number 1: 0,5 &mu;L arabinose, number 2: 1,0 &mu;L arabinose.<br>
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The rest is done just like earlier experiments with a total of 20 tubes because everthing is done twice.<br>
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<br>
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<strong>Results</strong><br>
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Some of the I1 (BBa_K2604001) JM109 and HB101 bacteria and the negative controls D1 and J1 (BBa_K2604003 and BBa_K2604000) have produced gas over the weekend.<br>
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We can not say for sure if the PBad promoter and the gas production enzyme pyruvate decarboxylase work. Fot that more gas production test have to be done. Also the negative controls produce gas. This is a problem. The JM109 and HB101 E.coli strains didn't produce gas in experiment .&nbsp;
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</div>
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<div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/c/c5/T--Rotterdam_HR--Exp5f1.jpeg');"></div>
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  <p>Figure 1: Gas testing before the weekend, II1.1 HB101 nr.1/  I1.2 HB101 nr.1/  I1.3, HB101 nr.1/  I1.1 JM109 nr.1
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</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/8/8c/T--Rotterdam_HR--exp5f2.jpeg');"></div>
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  <p>Figure 2: Gas testing after the weekend, II1.1 HB101 nr.1/  I1.2 HB101 nr.1/  I1.3, HB101 nr.1/  I1.1 JM109 nr.1</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/b/b1/T--Rotterdam_HR--exp5f3.jpeg');"></div>
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  <p>Figure 3: Gas testing before the weekend, I1.3 JM109 nr.1/ I1.2 JM109 nr.1/ J1 JM109 nr.1/ D1 HB101 nr.1</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/1/19/T--Rotterdam_HR--exp5f4.jpeg');"></div>
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  <p>Figure 4: Gas testing after the weekend, I1.3 JM109 nr.1/ I1.2 JM109 nr.1/ J1 JM109 nr.1/ D1 HB101 nr.1</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/d/dd/T--Rotterdam_HR--Exp5f5.jpeg');"></div>
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  <p>Figure 5: Gas testing before the weekend, D1 JM109 nr.1/ J1 HB101 nr.1</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/5/51/T--Rotterdam_HR--exp5f6.jpeg');"></div>
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  <p>Figure 6: Gas testing after the weekend, D1 JM109 nr.1/ J1 HB101 nr.1</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/d/df/T--Rotterdam_HR--exp5f7.jpeg');"></div>
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  <p>Figure 7: Gas production before the weekend, I1.3 JM109 nr.2/ I1.2 JM109 nr.2/ I1.1 JM109 nr.2/ I1.3 HB101 nr.2</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/c/c1/T--Rotterdam_HR--exp5f8.jpeg');"></div>
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  <p>Figure 8: Gas production after the weekend, I1.3 JM109 nr.2/ I1.2 JM109 nr.2/ I1.1 JM109 nr.2/ I1.3 HB101 nr.2</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/9/91/T--Rotterdam_HR--exp5f9.jpeg');"></div>
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  <p>Figure 9: Gas production before the weekend, D1 JM109 nr.2/ D1 HB101 nr.2/ J1 HB101 nr.2/ J1 JM109 nr.2 </p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/b/b6/T--Rotterdam_HR--exp5f10.jpeg');"></div>
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  <p>Figure 10: Gas production after the weekend, D1 JM109 nr.2/ D1 HB101 nr.2/ J1 HB101 nr.2/ J1 JM109 nr.2</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/7/7f/T--Rotterdam_HR--exp5f11.jpeg');"></div>
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  <p>Figure 11: Gas production before the weekend, I1.2 HB101 nr.2/ I1.1 HB101 nr.2</p>
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</div>
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<div class="notebook-image">
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  <div class="image-holder" style="background-image: url('https://static.igem.org/mediawiki/2018/3/31/T--Rotterdam_HR--exp5f12.jpeg');"></div>
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  <p>Figure 12: Gas production after the weekend, I1.2 HB101 nr.2/ I1.1 HB101 nr.2</p>
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</div>
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</div>
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</div>
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<div class="cardform">
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<div class="number"><b>06</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Testing gas production</h3>
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<label for="switch-140" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">October 11</h2>
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<h1 class="title">Assessing different ATP sensors</h1>
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<h3 class="date">October 15</h3>
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<label class="back" for="switch-141">&times;</label>
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<div class="attendees">Loraine Nelson | Elise Grootscholten | Paul Reusink</div>
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<div class="description">
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<p>Biobricks from iGEM were transformed into NEB10Beta. A few of these are sensors that react to ATP&nbsp;and can be used to detect living cells.&nbsp;The Bacteria were plated on agar plates with the right antibiotics to select the bacteria with the right plasmid in them. Every transformation was successful, since there were over 300 colonies on every plate. These colonies could than be used for further experiments and can be easily used for DNA isolation.&nbsp;</p>
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<p class="experiment"></p>
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<p><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px; word-wrap: break-word;">Materials:&nbsp; </span></p>
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<p style="background-color: transparent; box-sizing: content-box; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; orphans: 2; padding-bottom: 0px; padding-left: 0px; padding-right: 0px; padding-top: 0px; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">- Biobricks&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Nuclease free water<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- LB- agar<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Variable Volume Pipettes&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Sterile pipetpoints<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Chemically competent cells ( NEB10Beta, 1x10^9)&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Heater (42&deg;C) &nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Incubator + shaker (37&deg;C and 250 rpm)&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Eppendorf tubes (1,5 and 2 ml)&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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- Stable outgrow medium for NEB10Beta or SOC- outgrow medium&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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Method:&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24&deg;C). After incubation the DNA is transferred to Eppendorf tubes&nbsp;and put on ice.&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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For the transformation 1 ul of the DNA was added&nbsp;to 50 ul&nbsp;chemically competent cells strain&nbsp;NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube&nbsp;in a heater (42&deg;C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes.&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37&deg;C and 250 rpm.&nbsp;&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37&deg;C.&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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Results:<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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Tabel 1 shows the results of the plates which were incubated.&nbsp;</p>
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<table style="margin: 0px; width: 427px; text-align: left; color: rgb(51, 51, 51); text-transform: none; text-indent: 0px; letter-spacing: normal; font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; text-decoration: none; word-spacing: 0px; white-space: normal; box-sizing: border-box; orphans: 2; -webkit-text-stroke-width: 0px; background-color: transparent;" border="1" cellspacing="1" cellpadding="1">
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<tbody style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Biobricks:&nbsp;</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">Antibiotics:&nbsp;</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">Dilution:&nbsp;</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;">Colonies:</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">CFU:&nbsp;</td>
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</tr>
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<tr style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Bba-K352002</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">Can&nbsp;</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x&nbsp;</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;">&gt;300</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
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</tr>
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<tr style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Bba-K352002</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">Can</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">Centrifuged</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
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</tr>
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<tr style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Bba-K352003</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">Can</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
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</tr>
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<tr style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Bba-K352003</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">Can</td>
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<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">Centrifuged</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">&nbsp;- &nbsp; &nbsp; &nbsp;</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352004</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352004</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352005</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352005</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352006</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352006</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352007</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352007</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352008</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352008</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352009</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K352009</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K1390001</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K1390001</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K1390003</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K1390003</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K1023003</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K1023003</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Can</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K284001</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">amp</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Bba-K284001</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">amp</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Centrifuged</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
<tr style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Positive controle: Bba- K1023003</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">None</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;"><span style="display: inline !important; float: none; background-color: transparent; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">&gt;300</span></td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">- &nbsp;&nbsp;</td>
 +
</tr>
 +
<tr style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 101.49px;">Negative controle: outgrow medium</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 61.34px;">none</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 80.85px;">0x</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 68.07px;">0</td>
 +
<td style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; width: 87.25px;">-</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
 +
<p style="background-color: transparent; box-sizing: content-box; color: rgb(51, 51, 51); font-family: Arial,Helvetica,sans-serif; font-size: 12px; font-style: normal; font-variant: normal; font-weight: 400; letter-spacing: normal; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px; orphans: 2; padding-bottom: 0px; padding-left: 0px; padding-right: 0px; padding-top: 0px; text-align: left; text-decoration: none; text-indent: 0px; text-transform: none; -webkit-text-stroke-width: 0px; white-space: normal; word-spacing: 0px;">Tabel 1: results from the transformation with the biobricks.&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
Showed in the Tabel, the transformation gave&nbsp; enough colonies on each plate, which can be used for further experiments. There is no contamination, as seen by the negative control.&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
Conclusion:&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
The transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4&deg;C and can be used for further experiments.&nbsp;<br style="box-sizing: content-box; margin-bottom: 0px; margin-left: 0px; margin-right: 0px; margin-top: 0px;">
 +
&nbsp;</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>04</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing different ATP sensors</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">October 15</h2>
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 +
</svg>
 +
<h1 class="title">Assessing different ATP sensors</h1>
 +
<h3 class="date">October 15</h3>
 +
<label class="back" for="switch-142">&times;</label>
 +
</div>
 +
<div class="attendees">Loraine Nelson | Elise Grootscholten | Paul Reusink</div>
 +
<div class="description">
 +
<p>The bacteria were mini prepped and than transformed into BL21 DE3. BL21 DE3 has a T7 promotor which is needed for expression of the biobricks. The transformation was successful and multiple colonies were formed on the agar plates with antibiotics. From this plates we grow a few colonies onto new plates for further experiments.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<h1><span style="font-size: 16px;">Transformation Protocol</span> <span style="font-size: 16px;">NEB</span><br>
 +
<span style="font-size: 14px;"><span style="color: rgb(0, 0, 0);">Overview</span></span></h1>
 +
 +
<p><span style="font-size: 14px;"><span style="color: rgb(0, 0, 0);"><span style="font-family: arial,helvetica,sans-serif;">Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.</span></span></span></p>
 +
 +
<section class="paragraph  ">
 +
<h2><span style="font-size: 14px;">Protocol</span></h2>
 +
 +
<ol>
 +
<li>Thaw competent cells on ice.</li>
 +
<li>Chill approximately 5 ng (2 &mu;l) of the ligation mixture in a 1.5 ml microcentrifuge tube.</li>
 +
<li>Add 50 &mu;l of competent cells to the DNA. Mix gently by pipetting up and down or flicking the tube 4 to 5 times to mix the cells and DNA. Do not vortex.</li>
 +
<li>Place the mixture on ice for 30 minutes. Do not mix.</li>
 +
<li>Heat shock at 42&deg;C for 30 seconds*. Do not mix.</li>
 +
<li>Add 950 &mu;l of room temperature media* to the tube.</li>
 +
<li>Place tube at 37&deg;C for 60 minutes. Shake vigorously (250 rpm) or rotate.</li>
 +
<li>Warm selection plates to 37&deg;C.</li>
 +
<li>Spread 50 to100 &mu;l of the cells and ligation mixture onto the plates.</li>
 +
<li>Incubate overnight at 37&deg;C.
 +
<p>* Please note: For the duration and temperature of the heat shock step as well as for the media to be used during the recovery period, please follow the recommendations provided by the competent cells� manufacturer.<br>
 +
<br>
 +
1 ul isolated K1023003 biobrick + 50 ul NEB10b&egrave;ta&nbsp;<br>
 +
Positive and negative control without antibiotics<br>
 +
<br>
 +
&nbsp;</p>
 +
</li>
 +
</ol>
 +
</section>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>05</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing different ATP sensors</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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 +
</svg>
 +
<h1 class="title">Assessing Gas Production BioBricks in E.Coli</h1>
 +
<h3 class="date">October 16</h3>
 +
<label class="back" for="switch-143">&times;</label>
 +
</div>
 +
<div class="attendees">Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Glycerolstocks have been made of several bacteria with plasmids we created or from the iGEM kit.</p>
 +
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>22</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing Gas Production BioBricks in E.Coli</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
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<h2 class="date" style="color: white;">October 16</h2>
 +
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<svg viewBox="0 0 24 24">
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Assessing CooA Production</h1>
 +
<h3 class="date">October 16</h3>
 +
<label class="back" for="switch-144">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin van der Meulen</div>
 +
<div class="description">
 +
<p>After performing a digestion on the 16th we ran a gel electrophoresis, from which we could conlcude that only the combinations of J23100+B0031, J23100+B0032 and K352001+B0015 seemed to be correct, though because of the small insert size in the promoter and rbs combinations it is difficult to say. From these combinations overnight cultures where made, which we prepped today.&nbsp;</p>
 +
 +
<table border="1" cellpadding="1" cellspacing="1" style="width: 500px;">
 +
<tbody>
 +
<tr>
 +
<td>J23100+B0031</td>
 +
<td>125,9 ng/uL</td>
 +
</tr>
 +
<tr>
 +
<td>J23100+B0032</td>
 +
<td>334,1 ng/&mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>K352001+B0015</td>
 +
<td>240,3 ng/&mu;L</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<br>
 +
After this we've discussed our progress with our PI's,&nbsp;
 +
<p class="experiment"></p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>08</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Assessing CooA Production</h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<h2 class="date" style="color: white;">October 16</h2>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
 +
</svg>
 +
<h1 class="title">Chemo competent cells </h1>
 +
<h3 class="date">October 16</h3>
 +
<label class="back" for="switch-145">&times;</label>
 +
</div>
 +
<div class="attendees">None</div>
 +
<div class="description">
 +
<p>New competent cells were made. This time we used the strains JM109 and HB101, since these strains don't produce gas of themselfes. These strains can be used for measurement on our gas production system. The cells are competent enough to use.&nbsp;</p>
 +
 +
<p class="experiment"></p>
 +
<p><span style="font-size:14px;"><strong>Experiment 2: Chemocompetent cells of E.coli strains&nbsp;JM109 and HB101</strong></span><br>
 +
28-09-18<br>
 +
<br>
 +
<strong>Methods</strong><br>
 +
The methods are for JM109 and HB101 both apart.<br>
 +
2 ul pUC19 + 50 ul competent cells<br>
 +
With ampicilin: not diluted, 10^-1, 10^-2, 10^-3, 10^-4<br>
 +
Without ampicillin: not diluted, 10^-4, 10^-5, 10^-6<br>
 +
Negative control: SOC medium.<br>
 +
<br>
 +
200 ul not diluted --&gt;<br>
 +
30 ul + 270 ul SOC (10^-1) --&gt;&nbsp;<br>
 +
30 ul + 270 ul SOC (10^-2) --&gt;&nbsp;<br>
 +
30 ul + 270 ul SOC (10^-3) --&gt;&nbsp;<br>
 +
30 ul + 270 ul SOC (10^-4) --&gt;&nbsp;<br>
 +
30 ul + 270 ul SOC (10^-5) --&gt;&nbsp;<br>
 +
30 ul + 270 ul SOC (10^-6)<br>
 +
<br>
 +
Results<br>
 +
JM109: 3,4*10^6 cfu/ug DNA<br>
 +
HB101: 4,4*10^6 cfu/ug DNA</p>
 +
 +
</div>
 +
</div>
 +
 +
<div class="cardform">
 +
<div class="number"><b>02</b></div>
 +
<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Chemo competent cells </h3>
 +
<svg viewBox="0 0 24 24">
 +
<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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</svg>
 +
<h1 class="title">Preparing DNA for Submission</h1>
 +
<h3 class="date">October 17</h3>
 +
<label class="back" for="switch-146">&times;</label>
 +
</div>
 +
<div class="attendees">Dustin Vermeulen | Elise Grootscholten | Randall de Waard</div>
 +
<div class="description">
 +
<p>Today we set our control digest from the 9th of october on gel (the gel was inconclusive because of poor quality), after which we put ~500 ng of DNA on our 96 wells submission plate (both the pSB1C3 and pSB1K3 (sequenced) versions of our biobricks). This was then dried in a 50&deg;C stove for around 2 hours (until all wells were drie).</p>
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<p class="experiment"></p>
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<div class="cardform">
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<div class="number"><b>04</b></div>
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<h3 class="title" style="color: white; padding: 10px 5px 5px 5px;">Preparing DNA for Submission</h3>
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<path d="M9.46,6.28L11.05,9C8.47,9.26 6.5,11.41 6.5,14A5,5 0 0,0 11.5,19C13.55,19 15.31,17.77 16.08,16H13.5V14H21.5V16H19.25C18.84,17.57 17.97,18.96 16.79,20H19.5V22H3.5V20H6.21C4.55,18.53 3.5,16.39 3.5,14C3.5,10.37 5.96,7.2 9.46,6.28M12.74,2.07L13.5,3.37L14.36,2.87L17.86,8.93L14.39,10.93L10.89,4.87L11.76,4.37L11,3.07L12.74,2.07Z"></path>
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<label for="switch-146" class="more-info">Show more</label>
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<h2 class="date" style="color: white;">October 17</h2>
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<div class="text-container relevant">
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  <h1>Relevant pages</h1>
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    <a href="https://2018.igem.org/Team:Rotterdam_HR/Results" class="nav-card">
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      <h2>Results</h2>
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<path d="M19,3H14.82C14.4,1.84 13.3,1 12,1C10.7,1 9.6,1.84 9.18,3H5A2,2 0 0,0 3,5V19A2,2 0 0,0 5,21H19A2,2 0 0,0 21,19V5A2,2 0 0,0 19,3M12,3A1,1 0 0,1 13,4A1,1 0 0,1 12,5A1,1 0 0,1 11,4A1,1 0 0,1 12,3M7,7H17V5H19V19H5V5H7V7M7.5,13.5L9,12L11,14L15.5,9.5L17,11L11,17L7.5,13.5Z" />
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      <h2>Hardware</h2>
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Latest revision as of 19:46, 17 October 2018

Notebook

Notebook

As a team with only one computer scientist (and one electrical engineer who can program), we wanted a notebook that automatically updates whenever someone adds something to our lab journal. Luckily, we gained a sponsor that gave use an electronic lab journal as sponsorship. With this, we can use their API to download specific sections from our lab journal and display them on or wiki. But, as a multidisciplinary team we have multiple kinds of notebook entries: Software, Hardware and Wetlab. Therefore we also used the Google Drive RESTful API for downloading software and hardware entries from google sheets files.

More info about the notebook generator
show wetlab entries
 show hardware entries
 show software entries

Team page

May 10

Rens Boeser
Created the first prototype for the team page

01

Team page

May 10

Footer

May 20

Rens Boeser
Social media created

01

Footer

May 20

Footer

May 21

Rens Boeser
Social media buttons same color and fixed iGEM style issues

02

Footer

May 21

Notebook

May 24

Rens Boeser
Multiple week summary styles created for the notes

01

Notebook

May 24

Header

May 27

Rens Boeser
Added the header with mobile compatibility als resized logos and readded them to the header

01

Header

May 27

Main template

May 28

Rens Boeser
Added template header and footer with 'under construction' content

01

Main template

May 28

Header

May 28

Rens Boeser
Small fixes and menu adjustments

02

Header

May 28

Team

May 29

Rens Boeser
Added a PI list with the secondary PI

02

Team

May 29

Main page

June 7

Rens Boeser
Designed and added content holders to the index Page

01

Main page

June 7

Team

June 9

Rens Boeser
Added myself to the teammember Page and did some bugfixing

03

Team

June 9

Header

June 9

Rens Boeser
Started working on a better looking menubar

03

Header

June 9

Header

June 10

Rens Boeser
Added the ability to add sub-menus to items in the menu bar. Integration into the website and added indication for which Page you are on

04

Header

June 10

Construction pages

June 13

Rens Boeser
Updated all Pages to the new style with an under construction sign

01

Construction pages

June 13

Header

June 14

Rens Boeser
Added a way to recognise Pages under construction using the menubar

05

Header

June 14

Main page

June 14

Rens Boeser
Updated the content containers so that they are responsive

02

Main page

June 14

Architecture design [TC]

June 18

Tom van Dijk | Mike Vrieswijk
The first draft of the architecture design for the temperature controller was made.

01

Architecture design [TC]

June 18

Human Practices page

June 22

Rens Boeser
Content added to the human practices page

01

Human Practices page

June 22

Main page

June 23

Rens Boeser
Content created and added to the main page

03

Main page

June 23

Architecture design [TC]

June 24

Tom van Dijk | Mike Vrieswijk
Today a few changes have been made to the architecture design of the temperature controller. The choice was made to use a premade power supply as opposed to a self-made one, because this was considering the time left to finish this device the best choice. A few alterations were made to make the architecture design better readable.

02

Architecture design [TC]

June 24

First prototype [TC]

June 25

Tom van Dijk | Mike Vrieswijk
A 3d drawing was made for the casing that will hold the temperature controller. The first prototype was printed, but was determent to be inadequate due to the lack of enough space for the cooler block.

01

First prototype [TC]

June 25

Team

June 26

Rens Boeser
Added the remaining instructors to the team

04

Team

June 26

Second prototype [TC]

June 26

Tom van Dijk | Mike Vrieswijk
In the drawing space was added to fit the cooler block for the temperature controller. Slits were added to the drawing for the nuts to slide into. The altered drawing was printed. The newly printed parts, cooler block, fan and Peltier element were assembled. Everything fit together.

02

Second prototype [TC]

June 26

Assessing CooA Production

June 26

Dustin van der Meulen
To start testing the production of CooA, we firstly resuspended and transformed the BioBricks from the kit witch we wanted to use (Table 1). This has been done using the iGEM Kit plate instructions (Protocol 1, step 1 - 3) and the NEB 10 béta transformation protocol adapted New England Biolabs (Protocol 2), using bought cells with a reported transformation efficienty of 1-3 10^9 cfu/μg pUC19. Inucbation was at 37°C.

Table 1 : Resuspended BioBricks from the iGEM 2018 Distribution kit and their uses.
BBa_K592009 Blue Chromoprotein AmilCP
BBa_B0031 Ribosomal binding site (strong), derived from BBa_B0030
BBa_B0032 Ribosomal binding site (medium), derived from BBa_B0030
BBa_B0030 Ribosomal binding site (weak)
BBa_B0015 Double terminator (BBa_B0010 & BBa_B0012)
BBa_J23100 Strong Constitutive Anderson Promotor
BBa_J23105 Medium Constitutive Anderson Promotor
BBa_J23113 Weak Constitutive Anderson Promotor
BBa_J45199 Banana odor enzyme (ATF1) generator
BBa_K1184000 Killer Red
BBa_K352001 CooA
BBa_K352011 CooA responsive system

After checking the plates on the 28th of June, we recieved the following results (Table 2):

Table 2 : Kolonies found after overnight growth.
BBa_K592009 20 colonies
BBa_B0031 4 colonies
BBa_B0032 5 colonies
BBa_B0030 1 colonies
BBa_B0015 6 colonies
BBa_J23100 1 colonies
BBa_J23105 98 colonies
BBa_J23113 19 colonies
BBa_J45199 0 colonies
BBa_K1184000 4 colonies
BBa_K352001 2 colonies
BBa_K352011 4 colonies

01

Assessing CooA Production

June 26

Testrun [TC]

June 27

Tom van Dijk | Mike Vrieswijk
A short test run was done with the newly assembled cooler. The test concluded that the temperature controller could cool a water droplet to the point of freezing and heat it back up to its boiling point.

03

Testrun [TC]

June 27

Assessing CooA Production

June 29

Dustin van der Meulen

After making a 20 mL overnight culture of our NEB 10 béta cells containing our BioBricks, we performed a mini prep (Protocol 3). Using our nanodrop spectrophotometer  which gave us the following DNA concentrations:
 

BBa_K592009 72,5 ng/μL
BBa_B0031 118,9 ng/μL
BBa_B0032 52,5 ng/μL
BBa_B0030 488,6 ng/μL
BBa_B0015 53,5 ng/μL
BBa_J23100 148,3 ng/μL
BBa_J23105 96,3 ng/μL
BBa_J23113 151,8 ng/μL
BBa_K118400 140,4 ng/μL
BBa_K352001 77,7 ng/μL
BBa_K352011 76,8 ng/μL

After assessign our results, in the future we will perform the ethanol carry-over steps.

Also from the previously mentioned overnight culture 1 mL has been used to make Glycerol Stock (Protocol 4)

02

Assessing CooA Production

June 29

Draft decision

July 2

Tom van Dijk | Mike Vrieswijk
The choice to use a computer PSU as a power supply was made. The choice was made on the grounds, that computer PSU have a standard 12, 5 and 3.3V output. This is beneficial because these are the voltages necessary for the selected components.

01

Draft decision

July 2

Assessing CooA Production

July 2

Dustin van der Meulen

Today we performed a mini prep, digestion, dephosphorilation and ligation (following protocols 4 and 5) of a few promotor and RBS BioBricks. Because of administrative complications only the variants with J23113 and a RBS have been digested correctly.

03

Assessing CooA Production

July 2

Assessing CooA Production

July 2

Dustin van der Meulen

Today we transformed the previously made three combinations (BBa_J23113 + BBa_B0031 / B0032 / B0030) into chemically competent NEb 10 béta E.coli cells using the corresponding NEB protocol.

04

Assessing CooA Production

July 2

Architecture design [TC]

July 5

Tom van Dijk | Mike Vrieswijk
Changes were made to the architecture design of the temperature controller. Different voltage regulators were selected to fit the newly chosen power supply. To better readability of the architecture design a legend was added.

03

Architecture design [TC]

July 5

Assessing CooA Production

July 5

Dustin van der Meulen

After performing a mini prep and NotI digest on the 5th of July, and a gel electrophoresis today, we concluded that the cloning wasn't succesful.
 

05

Assessing CooA Production

July 5

Electrical circuit design [TC]

July 8

Tom van Dijk | Mike Vrieswijk
Custom footprints were made for multiple components and added to the parts library in the PCB design software.

01

Electrical circuit design [TC]

July 8

Electrical circuit design [TC]

July 9

Tom van Dijk | Mike Vrieswijk
The provisional electronic circuit design for the temperature controller was made.

02

Electrical circuit design [TC]

July 9

Assessing CooA Production

July 10

Dustin van der Meulen

Today we again performed a mini prep (overnight cultures from 10th of July) and digest. Sadly most digests did not give the expected results. Even though we had already progressed with the ligation. For now we've decided to progress with these ligation products.

 

06

Assessing CooA Production

July 10

Electrical circuit design [TC]

July 11

Tom van Dijk | Mike Vrieswijk
The electronic circuit design for the temperature controller was finished. Measuring point were added to the electronic circuit design.

03

Electrical circuit design [TC]

July 11

Stock up

July 11

Elise Grootscholten | Loraine Nelson | Paul Reusink
To have some basic biobricks ready, we stocked them by doing a plasmid isolation and freeze the DNA.

01

Stock up

July 11

Basic parts

July 11

Elise Grootscholten | Loraine Nelson | Paul Reusink
To start on a basic construct we did a digestion and ligation with the parts we isolated earlier. To prevent original constructs we did a defosphorylation after the digestion. We controlled the digestion by doing a gelelktroforeses.

01

Basic parts

July 11

Board layout [TC]

July 12

Tom van Dijk | Mike Vrieswijk
Started working on the board layout for the PCB of the temperature controller.

01

Board layout [TC]

July 12

Basic parts

July 12

Tom van Dijk | Randall de Waard
We continued with the ligated DNA by transformating it to Neb10Beta. This was plated onto agarplates and incubated for 1 day.

02

Basic parts

July 12

Board layout [TC]

July 13

Tom van Dijk | Mike Vrieswijk
Further work was done to the board layout for the temperature controller.

02

Board layout [TC]

July 13

Basic parts

July 13

Elise Grootscholten | Randall de Waard
The results of 7/12/2018 were collected, in action of this we decided to plate some ligated and transferred DNA from 7/12/2018 again. This we did because there wasn't a clear grow of colonies on the plates.

03

Basic parts

July 13

Blue White screening on paper

July 13

Elise Grootscholten | Randall de Waard
We tried to let bacteria grow on paper, therefore we used our own buisnesscards. We let the buisnesscards absorb some LB-medium and put them inside petridishes.

01

Blue White screening on paper

July 13

Board layout [TC]

July 15

Tom van Dijk | Mike Vrieswijk
Work has been done on the board layout for the temperature controller. The board layout is now finished.

03

Board layout [TC]

July 15

Assessing CooA Production

July 15

Dustin van der Meulen
After we transformed our ligation results from the 11th on the 12th, we recieved the following results on the 13th:


B0030+J23100: No growth
B0030+J23105: Unsure (small sports, maybe colonies)
B0031+J23100: Growth
B0031+J23105: Unsure (small sports, maybe colonies)
B0032+J23100: Growth
B0032+J23105: Unsure (small sports, maybe colonies)
K352001+B0015: Growth

On the 13th we retransformed the 4 ligations of which we didn't get conclusive results, which we looked at today:
B0030+J23100: No growth

B0030+J23105: Again unsure
B0031+J23105: No growth
B0032+J23105: No growth

After assessing the results we made overnight cultures of the following:

B0031+J23100

K352001+B0015

B0032+J23100

B0030+J23105 (1)

B0030+J23105 (2)

B0032+J23105

Also we retransformed the colonies we retransformed on the 13th again, and quickly spun our cells in a centrifuge, removing most of the supernatant and resuspending our cells before we spread them on Chloramphenicol plates.

07

Assessing CooA Production

July 15

Basic parts

July 16

Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Randall de Waard
To know if the ligation was succesfull we will controll the DNA, therefor we ented the bacteria's with DNA in LB-medium to grow overnight. We also plated the used bacteria's onto new plates to use for further experiments.

04

Basic parts

July 16

Blue White screening on paper

July 16

Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Randall de Waard
We did a blue white screening on our buisnesscards to see if bacteria had grew on them. We transferred the buisnesscards to new petrydishes and added ITPG and X-gal to the petrydishes.

02

Blue White screening on paper

July 16

Notebook

July 17

Rens Boeser
Frontend design for the notebook

02

Notebook

July 17

Basic parts

July 17

Rens Boeser | Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Randall de Waard
The resulst of 7/16/2018 showed that the DNA didnt ligate as planned. To be able to digest enough DNA we did a plasmid isolation again. We digested this DNA and froze it for further use.

05

Basic parts

July 17

Stock up

July 18

Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Mike Vrieswijk | Randall de Waard
We did another round of plasmid isolation for stocking.

02

Stock up

July 18

Notebook

July 19

Rens Boeser
Designed a way to follow experiments that take up multiple days and eventually designed a notebook entry page and made it a responsive page

03

Notebook

July 19

Basic parts

July 19

Dustin van der Meulen | Mei Ju Goemans| Randall de Waard
We digested all DNA we have in stock to test for our basic construct. We tested the DNA by doing a gelelktroforeses.

06

Basic parts

July 19

Notebook

July 21

Rens Boeser
Created a one page notebook. Made the notebook entry page more suited for the entry length

04

Notebook

July 21

Assessing Gas Production BioBricks in E.Coli

July 23

Elise Grootscholten | Randall de Waard

Transformations of the biobricks K133071, K173003 and I13453 are performed. K133071 will produce CO2 if there's pyruvate present and K173003 will produce CO2 if urea is present. I13453 is a promotor which will work if there's arabinose present.                                                                                                        
The biobricks were transformed first into electrocometent cells and later into chemical competent cells from the strain NEb10Beta. After transformations the culture was plated on agar plates with antibiotics. The first transformations didn't work out, but the second did, because then the right competent cells were used. 

Experiment 1: Transformation biobricks

Transformation of the biobricks K133071, K173003 and I13453 in NEB10Bèta.

Materials 

- Biobricks (Bba_K133071, Bba_K173003 and Bba_I3453) 
- Nuclease free water
- LB- agar
- Variable Volume Pipettes 
- Sterile pipetpoints
- Chemically competent cells ( NEB10Beta, 1x10^9) 
- Heater (42°C)  
- Incubator + shaker (37°C and 250 rpm) 
- Eppendorf tubes (1,5 and 2 ml) 
- Stable outgrow medium for NEB10Beta or SOC- outgrow medium 
-Chloramphenicol (40 mg/mL)

Methods
-K173003 --> pSB1C3 backbone, plate 6, well 15N
-K133071 --> pSB1C3 backbone, plate 6, wel 21A
-I13453 --> pSB1C3 backbone, plate 3, well 19O

To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24°C). After incubation the DNA is transferred to Eppendorf tubes and put on ice. 

For the transformation 1 ul of the DNA was added to 50 ul chemically competent cells strain NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube in a heater (42°C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes. 
For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37°C and 250 rpm.  

After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37°C. 

Results
The first transformation was done in electrocompetent cells instead of chemocompetent cells so the first transformation did not succeed.

 

Conclusion

The second transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4°C and can be used for further experiments. 
The CFU from the negative controle will be plated on a plate with antibiotics to make sure there isn't a plasmid in the bacteria. The outgrow medium will be sterilized by filter again. 
 

01

Assessing Gas Production BioBricks in E.Coli

July 23

ATP sensor

July 24

Rens Boeser | Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Randall de Waard
New biobricks, used to make an ATP sensor, were transformed into NEB10Beta. The culture was than plated onto agar and incubated for 1 day.

01

ATP sensor

July 24

Gas output

July 24

Rens Boeser | Mei Ju Goemans | Elise Grootscholten | Dustin van der Meulen | Loraine Nelson | Suzanne Romeijn | Randall de Waard
New biobricks, which produces gasses, were transformed into NEB10Beta. The culture was than plated onto agar and incubated for 1 day.

01

Gas output

July 24

Notebook generator

July 24

Rens Boeser
Started working on an automatic notebook creator

01

Notebook generator

July 24

Assessing Gas Production BioBricks in E.Coli

July 24

Elise Grootscholten | Randall de Waard

To be able to always get the necessary biobricks, there have been made glycerolstocks of the transformed biobricks K133071, K173003 and I13453. The glycerolstocks are stored at -80 degrees Celsius. If needed, they can be retrieved from this storage to use for experiments. 

Experiment 2: Glycerolstocks

Making glycerol stocks of the biobricks K133071, K173003, I13453. 

Materials
- 87% glycerol 
- culture with biobrick plasmids
- LB-medium
- Variable Volume Pipette
- Sterile pipetpoints

Methods
The colonies from the transformation were used to make an overnight culture. The culture was made by putting a colony in LB-medium and then the culture was incubated for a night at 37°C.  

The next day the cultures had grown close and were ready for a glycerol stock. For making glycerol stocks there was added 250 ul 87% glycerol to 1 ml culture. This was frozen in -80°C.  

The stocks were stored at -80°C in drawer 3, tower 1, drawer 

02

Assessing Gas Production BioBricks in E.Coli

July 24

Assessing different ATP sensors

July 24

Loraine Nelson | Elise Grootscholten | Paul Reusink

Today we did a transformation with new biobricks. The biobricks were built into chemically competent cells from the strain NEb10Beta. After transformation the culture was plated on agar plates with antibiotics. The results were checked the next day. 

Materials: 

- Biobricks (Bba-K1499004 and Bba- K1614019) 
- Nuclease free water
- LB- agar
- Variable Volume Pipettes 
- Sterile pipetpoints
- Chemically competent cells ( NEB10Beta, 1x10^9) 
- Heater (42°C)  
- Incubator + shaker (37°C and 250 rpm) 
- Eppendorf tubes (1,5 and 2 ml) 
- Stable outgrow medium for NEB10Beta or SOC- outgrow medium 

Method: 

To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24°C). After incubation the DNA is transferred to Eppendorf tubes and put on ice. 

For the transformation 1 ul of the DNA was added to 50 ul chemically competent cells strain NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube in a heater (42°C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes. 
For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37°C and 250 rpm.  

After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37°C. 

Results:
Tabel 1 shows the results of the plates which were incubated. 

Biobricks:  Antibiotics:  Dilution:  Colonies: CFU: 
Bba-K1614019  Can  0x  109 1*10^3 /ml
Bba-K1614019 Can Centrifuged >300    -
Bba-K1499004     Can 0x 33 3*10^2 /ml
Bba-K1499004         Can Centrifuged 228    -         
Positive controle: Bba- K1499004 None 0x > 300    -
Negative controle: outgrow medium none 0x 18    - 

Tabel 1: results from the transformation with the biobricks Bba-K1614019 and Bba-K1499004. 

The plates show CFU which can be used for further experiments.  The negative controle also has CFU, this shows that the outgrow medium was infected and not sterile. 
The plates are stored by 4°C. 

Conclusion: 

The transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4°C and can be used for further experiments. 
The CFU from the negative controle will be plated on a plate with antibiotics to make sure there isn't a plasmid in the bacteria. The outgrow medium will be sterilized by filter again. 

 

01

Assessing different ATP sensors

July 24

Notebook generator

July 25

Rens Boeser
Automatic generation of entries completed

02

Notebook generator

July 25

Google Drive API

July 25

Rens Boeser
Used the Google drive REST API to download the entries from drive

01

Google Drive API

July 25

Automatic uploader

July 25

Rens Boeser
Used the igemwiki API to update pages using software

01

Automatic uploader

July 25

Assessing Gas Production BioBricks in E.Coli

July 27

Elise Grootscholten | Randall de Waard

For further experiments there is isolated DNA needed of the biobricks J23100, K133071, K173003 and I134353. The DNA is isolated out of the bacteria with the help of a plasmid purifaction kit. After isolation this DNA can be used for digestion and ligation or other experiments. J23100 (from the glycerolstock): 325,59 ng/ul, K173003: 217,06 ng/ul, K133071: 186,79 ng/ul, I13453: 88,18 ng/ul

Experiment 3: Minipreps (DNA isolation)
27-07-18

Minipreps of the following biobricks in NEB10Bèta:
-J23100: Constitutive promoter
-K133071: Urea --> ammonia + CO2
-K173003: Pyruvate --> acetaldehyde + CO2
-I134353: Promoter (AraC protein binds with arabinose)

Materials 
- Mini prep, Plasmid Purification Kit, Machery Nagel 
- Eppendorf cups
- Variable Volume Pipets
- sterile pipet points
-Chloramphenicol (40 mg/mL)

Methods 
Miniprep protocol Plasmid Purification Kit, Machery Nagel
After the transformation we made an overnight culture from the colonies. The colonies were anted into 20 ml LB and incubated overnight at 37°C and 150 rpm. The culture was mini prepped the next day. For isolation about 6 mL overnight culture was used.


Plasmid DNA purification (NucleoSpin� Plasmid EasyPure)
Machery Nagel Mini preps
 

1 Cultivate

and harvest bacterial cells

12,000 x g, 30 s
 

2 Cell lysis 150 μL Buffer A1

250 μL Buffer A2

RT, up to 2 min

350 μL Buffer A3


3 Clarification of the lysate

> 12,000 x g, 3 min


4 Bind DNA

Load supernatant

1,000�2,000 x g, 30 s


5 Wash and dry silica

membrane

450 μL Buffer AQ

> 12,000 x g, 1 min


6 Elute DNA 50 μL Buffer AE

RT, 1 min

> 12,000 x g, 1 min


Results
J23100: 325,59 ng/ul
K173003: 217,06 ng/ul
K133071: 186,79 ng/ul
I13453: 88,18 ng/ul

Figure 1: Minipreps 27-07-18

03

Assessing Gas Production BioBricks in E.Coli

July 27

Assessing different ATP sensors

July 27

Loraine Nelson | Elise Grootscholten | Paul Reusink

To be able to always get the necessary biobricks, we made an glycerol stock after every transformation. The glycerol stocks are stored at -80 degrees Celsius. If needed, they can retrieved from this storage to use for experiments. 

Making glycerol stocks of the biobricks. 

Materials: 
- 87% glycerol 
- culture with biobrick plasmids
- LB-medium
- Variable Volume Pipette
- Sterile pipetpoints


Method: 

The colonies from the transformation were used to make an overnight culture. The culture was made by putting a colony in LB-medium and then the culture was incubated for a night at 37°C.  

The next day the cultures had grown close and were ready for a glycerol stock. For making glycerol stocks there was added 250 ul 87% glycerol to 1 ml culture. This was frozen in -80°C.  

The stocks were stored at -80°C in drawer 3, tower 1, drawer 

02

Assessing different ATP sensors

July 27

Automation program

July 29

Rens Boeser
Connected the page uploader to the notebook generator

01

Automation program

July 29

Automatic uploader

July 29

Rens Boeser
Created a program that uploads all updated pages

02

Automatic uploader

July 29

Assessing CooA Production

July 29

Dustin van der Meulen
After considering multiple tactics, we've decided to follow the standard iGEM 3A assembly in our following cloning steps. To this end we started with an overnight digest of all the different parts.
BioBrick BBa_ code Restriction enzyme 1 Restriction enzyme 2
J23100 EcoRI SpeI
J23105 EcoRI SpeI
J23113 EcoRI SpeI
B0030 EcoRI XbaI
B0031 EcoRI XbaI
B0032 EcoRI XbaI
K352001 SpeI PstI
B0015 XbaI PstI

Used pipette scheme from protocol 7 to prepare the DNA to be digested with multiple combinations of EcoRI-HF, SpeI-HF, PstI and XbaI.

With this we started a digest of the following BioBricks using the following Restriction enzyme couples:
BioBrick BBa_ code Restriction enzyme 1 Restriction enzyme 2
J23100 EcoRI SpeI
J23105 EcoRI SpeI
J23113 EcoRI SpeI
B0030 EcoRI XbaI
B0031 EcoRI XbaI
B0032 EcoRI XbaI
K352001 SpeI PstI
B0015 XbaI PstI

Digestion was started on the 30th of July, ~16:00, 37°C and 185 RPM.
Digestion was stopped on the 31st of July, ~10:00.
09

Assessing CooA Production

July 29

Making competent NEB10beta cells

July 30

Loraine Nelson | Jos Veldscholte

Made stock solution of MgSO4 and pure culture of NEB10 beta cells

Making 1 M sterile MgSO4
≈ 14.00: Dissolved 12,32 g MgSO4 in 50 mL demi water en filtered through 0.22 μm filter.

Making pure culture of NEB10 beta cells
≈15.00: 20 mL LB agar + 10 μL tetracyclin (50 mg/mL) to pour 1 plate.
16:30: streaked bacteria from glycerol stock: -80, section III, tower 1, shelf IV, box III, 6B
Left in 37°C room for o.n. growth.

01

Making competent NEB10beta cells

July 30

Page generator

July 31

Rens Boeser
Built a page builder that automatically adds the header and footer before uploading

01

Page generator

July 31

Google Drive API

July 31

Rens Boeser
Redone the google API handler for reusability and readability and made the google API handler generic

02

Google Drive API

July 31

Automation program

July 31

Rens Boeser
revamped the main program

02

Automation program

July 31

Assessing CooA Production

July 31

Dustin van der Meulen

Though we had started with our ligation using the rAPId dephos and ligation kit, we'd simultaniously put our DNA on gel, later revealing the digest to not have been succesful. Carefull examination of our work made us realise the used restriction enzyme dilutions to not have been made correctly.

Using the rAPid Dephos and Ligation Kit we performed a dephosphorilation and ligation of our DNA.
After checking our gel with our digestion products we later determined that the digestion didn't go as expected
 

10

Assessing CooA Production

July 31

Making competent NEB10beta cells

July 31

Loraine Nelson | Jos Veldscholte

Made Ksi broth (for competent cells) and started o.n. cultures

Make Ksi broth (for competent cells)
Dissolved 20 g Tryptone (Lab M limited) and 5 g Yeast extract (Lab M limited) in 1 liter demi water, added 10 ml 1 M NaCl and adjusted pH to 7.5 with KOH.
Autoclaved.
16:30
2 tubes with 2.5 mL Ksi inoculated with 1 colony NEB10beta from yesterday's plate. (Added 50 microliters of 1 M MgSO4 at 16:30 becauce I forgot to ad it to de autoclaved medium. 
Added 10 mL of 1 M MgSO4 to each half liter autoclaved medium (=Ksi).

02

Making competent NEB10beta cells

July 31

Automation program

August 1

Rens Boeser
Finished redoing the application for generating and uploading pages

03

Automation program

August 1

Making competent NEB10beta cells

August 1

Loraine Nelson | Jos Veldscholte

Made large NEB10 beta culture and aliquoted in 50 microliter portions and snap froze in dry ice/isopropanol

Making competent NEB10 beta cells
8:45:
Both cultures were fine.
Transfered one culture completely to 100 ml Erlenmeyer with 25 mL Ksi (RT). In shaking incubator at 37° C at 200 rpm.
Filled three one liter Erlenmeyers with 175 mL Ksi and put in shaking incubator in 37° C room to adjust temperature.
25 ml culture: at 8:40 OD600 was 0.64.
Transferred 8 ml to each 175 mL Erlenmeyer. Incubate at  37° C shaking rigorously (no speed indicator).
9:40: OD600 = 0.-9
10:30: OD600 = 0.30
10:50 OD600 = 0.46
10:58 OD600 = 0.55
Tranferred all cultures to 10 50 mL tubes and tare in pairs.
On ice 11:15
11:35: in Sorvall 4500 rpm, 15 min 0° C.
After the run, fluid was on the rotor and in the rotor holes.
Cleaned with lab desinfectant. Rinsed with water and dried with tissue.
From here on everything in the cold room with tubes on ice:
Resuspended pellets in 16,6 ml ice cold TFB1 (protocol "competente-cellen-maken"l) each with 10 mL plastic pipet and pipetboy. 
Combined into four 50 mL tubes. Centrifuged 15 min at 4500 rpm.
Resuspended pellets in 60 mL TFB1, kept on ice for 20 min and centrifuged 15 min 4500 rpm.
Respuspended pellets in 10,5 ml TFB2 (protocol "competente-cellen-maken"l). Left on Ice for 10 min.
Aliquotted into 50 μL portions in 2 mL conical tubes and snap froze in isopropanol on dry ice.
Transferred to -80°C Freezer.

03

Making competent NEB10beta cells

August 1

Notebook

August 2

Rens Boeser
Added month categorization to the page

05

Notebook

August 2

Assessing CooA Production

August 2

Dustin van der Meulen

We've redone the work mentioned in the previous CooA entry, though today our gel electrophoresis revealed we were only partially succesful, as only the digests with EcoRI and SpeI seem to be correct.

Experiment 9 and 10 have been repeated with a new enzyme dilution.
7th of august: review of the digestions on gel reveal that only the digestions with EcoRI-HF and SpeI-HF worked, while the PstI and XbaI digestion did not retrieve the expected results

11

Assessing CooA Production

August 2

Making competent NEB10beta cells

August 2

Loraine Nelson | Jos Veldscholte

Tested competence by transformation of pUC19

Test sample from NEB competent cells contains 50 pg pUC19 per μL.
Used 2 μL per transformation (= 100 ng).
Took three tubes from one of the two boxes.
A 100 ng pUC19
B 100 ng pUC19
C nothing
30 min on ice (11:00)
A 30 s heatshock in waterbath (11:30)
B 90 s heatshock in waterbath (11:30)
C was kept on ice still.
5 min on ice.
Added 1 ml of LB (11:40)
60 min shaking in 37 °C room (shaking slowly)

Poured LB agar plates with 50μg/ml Ampicillin and one plate without antibiotic.

12:40
Added 1 μL of culture A or B to 99 μL LB (A4 and B4)
Added 10 μL of culture A or B to 90 μL LB (A3 and C3)
Plated A3 and 4 and C3 and 4 on LB agar amp.
Plated 100 μL A, B and C (A2, B2 and C2) on LB agar amp.
Plated 100 μL C on plate without antibiotics (to test whether cells are alive)
Centrifuged A and B 6000 rpm in microcentrifuge for 2 min.
Aspirated supernatant.
Vortexed bacteria in remainder of the medium (>100 μL)
Plated on LB agar amp. plates (A1 and B1).

Consequently:
A was heat shocked for 30 s.
B was heat shocked for 90 s.
1 = 90% of the transformation, 2 = 10%, 3 = 1% and 4 = 0,1%

Put in automated incubator. Not sure about whether the sytem wil work. Is it on?
Colonies to be counted next monday by Dustin.
If there are no colonies visible, please extend incubation time!

04

Making competent NEB10beta cells

August 2

Making competent NEB10beta cells

August 3

Loraine Nelson | Jos Veldscholte

Counted colonies. Result: no transformation.

Colonies where counted at ~10:00. Only plate A1 showed signs of colonies (~15). All the others where empty.
After leaving the cells in the incubator for an extra half a day, there where no new colonies.

05

Making competent NEB10beta cells

August 3

Assessing CooA Production

August 4

Dustin van der Meulen
The previous two CooA entries have been repeated, though this time we remade the PstI and XbaI enzyme dilutions. This time the digests where as expected.

Experiment 11 has been repeated, but with new dilutions of PstI and XbaI.
After the incubation it was noted that the cup containging BBa_B0015 was almost empty, so it will have to be digested again. (did not happen) --> in the end we had enough

13th of august --> dephos and ligation was peformed on the obtained biobrick combinations following protocol 8
 

12

Assessing CooA Production

August 4

Making competent NEB10beta cells

August 7

Loraine Nelson | Jos Veldscholte

Repeated transformation.

Part 4 has been repeated, repeating the same steps but adding two extra positive controlls.

06

Making competent NEB10beta cells

August 7

Assessing different ATP sensors

August 9

Loraine Nelson | Elise Grootscholten | Paul Reusink

For further experiments we needed isolated DNA. The DNA is isolated out of the bacteria with the help of a plasmid purifaction kit. After isolation this DNA can be used for digestion and ligation or other experiments.

Materials: 
- Mini prep, Plasmid Purification Kit, Machery Michels 
- Eppendorf cups
- Variable Volume Pipets
- sterile pipet points


Method: 
After the transformation we made an overnight culture from the colonies. The colonies were anted into 20 ml LB and incubated overnight at 37°C and 150 rpm. The culture was mini prepped the next day. For isolation, 5,4 ml was added step wise in a 2ml Eppendorf cup. The culture was then centrifuged at >12000x g for 30 seconds. The supernatant was deposed, and the pellet was resupended with A1 buffer and vortexed. To the resuspended culture was then. 

03

Assessing different ATP sensors

August 9

Assessing Gas Production BioBricks in E.Coli

August 10

Elise Grootscholten | Randall de Waard

The biobricks J23100, K133071, K173003 and I134353 were sucessfully digested after the second time. After the digestions the biobricks K133071 and K173003 were dephosphorylated and ligated with the inserts J23100 and I13453. This was done in the original backbone of K133071 and K173003, and not another control backbone. To know if the biobricks were right ligated this was done by testing practically. See the experiments: Testing gas production. 

Experiment 4: Digestion, Defosforylation and ligation
Digestions
30-07-18 and 08-08-18

Biobricks: J23100, K133071, K173003 and I13453

Used pipette scheme (Table 1) to prepare the DNA to be cut with multiple combinations of EcoRI-HF, SpeI-HF, PstI and XbaI.

DNA ~ 2 μg
NEBbuffer 2.1 5 μL
Restriction enzyme 1 (1 U/μL) 2 μL
Restriction enzyme 2 (1 U/μL) 2 μL
Nuclease free water Fill to 100 μL
(Table 1 ; Double Digest Pipette Scheme using : https://nebcloner.neb.com/#!/redigest, and doubling the amount)


Biobricks:
J23100: Constitutive promotor
K133071: Urea --> ammonia + CO2
K173003: Pyruvate --> acetaldehyde + CO2
I134353: Promotor (AraC protein binds with arabinose)

With this we started a digest of the following BioBricks using the following Restriction enzyme couples (Table 2)
 
Number BioBrick BBa_ code Restriction enzyme 1 Restriction enzyme 2
9 J23100 (35 bp) EcoRI SpeI
11 K133071 (1707 bp) EcoRI XbaI
12 K173003 (3052 bp) EcoRI XbaI
10 I13453 (130 bp) EcoRI SpeI

Table 2: Biobricks and restriction enzymes

Digestion was started on the 30th of July, ~16:00, 37°C and 185 RPM.
Digestion was stopped on the 31st of July, ~10:00.

This has been done in tandem with the corresponding part from Cloning Mulitple BioBricks to Assess CooA Production Part X 



Dephosphorylation
10-08-18

K133071 and K173003 were dephosphoralized:
 

Volume Compound
25 μL Vector DNA (0,5 μg)
3 μL phosphatase buffer (10x)
1 μL milli Q
1 μL phosphatase (1 U/μL)
30 μL Total
Table 3: Pipetting scheme

The new DNA concentration of the reaction mix is 0,5 μg/30 μL
The reaction mix was incubated for 10 minutes at 37 °C
The phosphatase was inactivated for 2 minutes at 75 °C
 
For the ligation mix 100 ng of vector DNA was used from the dephosphorylation reaction mix.
The following vector-insert combinations were ligated:
Vector Insert 1 Insert 2
K133071 J23100 I13453
K173003 J23100 I13453
Table 4: Biobrick combination

The original plasmids of the vector and inserts that were digested are about the same size in bp and the ratio vector DNA-insert DNA is determined to be 1:3. Therefore, the amount of insert DNA that was used for the ligation was 140 ng (rounded up to 7 μL of the digestion reaction mix). Because there was only 5 μL left of the K352001 insert DNA, the total 5 μL were used and an extra 2 μL of milli Q water were added to the pre-ligation mix.

Pre-Ligationmix:
Volume Compound
6 μL Vector DNA (0,5 μg/30 μL)
7 μL Insert DNA (2 μg/100 μL)
4 μL Delutionbuffer (5x)
3 μL Milli Q
20 μL Total
Table 5: Pre-ligationmix pipette scheme

Ligationmix:
incubation at room-temperature for 20 minutes.
Volume Compound
10 μL Pre-ligationmix
2 μL 10x T4 ligationbuffer
1 μL T4 DNA ligase (1U/μL)
8 μL Milli Q
20 μL Total
Table 6: Ligationmix pipette scheme
 
All the vectors contain the CamR gene. 30 plates were poured with Cam (14 for the transformed cells (100μL), 14 for the transformed cells (900 μL centrifuged and resuspended in 100 μL and 1 positive and 1 negative control)

 

04

Assessing Gas Production BioBricks in E.Coli

August 10

Urea and sodium pyruvate test for resistance E.coli

August 10

Randall de Waard | Elise Grootscholten

Testing different amounts of urea and sodiumpyruvate to know which concentrations the bacteria survive.

Experiment 1: Testing different concentrations of Urea and Sodium pyruvate
08-08-18

Testing different amountsof urea and sodiumpyruvate to know which concentrations the bacteria survive.

-First let the LB agarose heat up (95 degrees Celcius)
-Second, weigh urea and sodium pyruvate in beakers (see table 1)

What How much Solution
Urea 1 g 1 mL purified water
Urea 2,4 g 2 mL purified water
Urea 4,8 g 4 mL purified water
Sodium pyruvate 5,7 mL None

Table 1: weighing urea and sodium pyruvate for LB agarose plates.
-Purify these substances, so it is free of bacteria. Work beneath the flame and pour it in to plastic 50 mL tubes.
-Pour the LB agarose in the plastic tubes when it's about 45 degrees Celcius and mix gently.
-Pour the LB agarose mix in a petridish and wait until it's dry. Do this for all the plastic tubes.
-Also fill 2 petridishes with LB agarose without other substances (positive and negative control).
-prick a BL21(DE3) bacteria and put it in a 100 mL erlenmeyer with 20 mL culture medium.

09-08-18

Pipette 100 ul overnight culture on each plate, except on the negative control, and divide it. (24h 37 degrees Celcius)

 

Results
 

What How much Solution Result
Urea 1 g 1 mL purified water Grow
Urea 2,4 g 2 mL purified water No grow
Urea 4,8 g 4 mL purified water No grow
Sodium pyruvate 5,7 mL - Grow
Positive control - - Grow
Negative control - - No grow

Table 1: Results experiment 1

Conclusion

More tests will be done with urea and sodiumpyruvate because with 1 g urea the bacteria live and with 2,4 g urea the bacteria die.
The bacteria survive 5,7 mL sodiumpyruvate so we will test with higher concentrations.
 

01

Urea and sodium pyruvate test for resistance E.coli

August 10

Assessing Gas Production BioBricks in E.Coli

August 13

Elise Grootscholten | Randall de Waard

Transformations of the biobricks (K133071 + J23100), (K13071 + I1345), (K173003 + J23100) and (K173003 + I13453) in NEB10Bèta. There was no grow except for the biobrick combination K133071 + J23100.

Experiment 5: Transformation

13-08-2018
Transformations of the ligations of experiment 4

Materials

-Chloramphenicol (40 mg/mL)
-LB-agar
-Ligations
-Heat block,42 degrees Celcius
-Shaker, 37 degrees Celsius 
-Chemo competent cells NEB10bèta (New England Biolabs)

Methods

10 ul ligationmix is used for every transformation

Transformation Protocol
Overview

Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.

Protocol

  1. Thaw competent cells on ice.
  2. Chill approximately 5 ng (2 μl) of the ligation mixture in a 1.5 ml microcentrifuge tube.
  3. Add 50 μl of competent cells to the DNA. Mix gently by pipetting up and down or flicking the tube 4�5 times to mix the cells and DNA. Do not vortex.
  4. Place the mixture on ice for 30 minutes. Do not mix.
  5. Heat shock at 42°C for 30 seconds*. Do not mix.
  6. Add 950 μl of room temperature media* to the tube.
  7. Place tube at 37°C for 60 minutes. Shake vigorously (250 rpm) or rotate.
  8. Warm selection plates to 37°C.
  9. Spread 50�100 μl of the cells and ligation mixture onto the plates.
  10. Incubate overnight at 37°C.

    * Please note: For the duration and temperature of the heat shock step as well as for the media to be used during the recovery period, please follow the recommendations provided by the competent cells� manufacturer.


Instead of 2 ul of the ligation mix, use 10 ul ligation mix.

05

Assessing Gas Production BioBricks in E.Coli

August 13

Assessing Gas Production BioBricks in E.Coli

August 14

Elise Grootscholten | Randall de Waard

A colony PCR is done for the NEB10Bèta E.coli cells with expected the biobrick combination of K133071 with J23100. Nevertheless, on a gel the difference with and without promotor couldn't be seen. So there must be another way of proving the right biobricks are there.

Expermiment 6: Colony PCR K133071 + J23100
14-08-18

Mastermix

What 1 reaction 30 reactions
Taq buffer(10x) 5 ul 150 ul
10 mM dntp 1 ul 30 ul
10 uM Forward primer 1 ul 30 ul
10 uM Reverse primer 1 ul 30 ul
Taq polymerase 0,25 ul 7,5 ul
Nucease free hydrogen 41,75 ul 1252,5 ul
Total 50 ul 1500 ul

 

06

Assessing Gas Production BioBricks in E.Coli

August 14

Assessing Gas Production BioBricks in E.Coli

August 15

Elise Grootscholten | Randall de Waard
The ligations and transformations of the biobricks K173003 + J23100, K173003 + I13453, and K133071 + I13453 have been performed again. There was a lot of grow on the agar plates. To know if the biobricks were right ligated this was done by testing practically. See the experiments: Testing gas production. 

Experiment 7: Ligation of biobricks
15-08-18

K173003 + J23100
K173003 + I13453
K133071 + I13453
 

Volume Compound
6 μL vector DNA (0,5 μg/30 μL) (100 ng)
14 μL insert DNA (2 μg/100 μL) (280 ng)
4 μL dilutionbuffer (5x)
1 μL milli Q water
25 μL Total


Use 10 mL

Ligationmix:

Volume Compound
10 μL pre-ligationmix 
2 μL 10x T4 ligationbuffer
1 μL T4 DNA ligase (1U/μL)
8 μL milli Q water
20 μL Total

incubation at room-temperature for 20 minutes.
Store at -20 degrees Celcius

07

Assessing Gas Production BioBricks in E.Coli

August 15

Assessing CooA Production

August 16

Dustin van der Meulen

We've performed a colony PCR on 15 random (though not the red/purple -esque colonies, which are collored only because they contain BBa_J04450) colonies from J23105+B0032 on the 13th. We also ligated the following biobricks anew:
B0031 + J23105
B0031 + J23113
B0032 + J23100
B0032 + J23105

We also transformed these biobricks into E.coli on the 16th, prepping and digesting them later on, though these and the PCR did not give us the result we'd hoped for.

Today a colony PCR will be performed to check our results from last friday, namely the transformation product of BBa_B0032 en BBa_J23105:

Mastermix

What 1 reaction 30 reactions
Taq buffer(10x) 5 ul 150 ul
10 mM dntp 1 ul 30 ul
10 uM Forward primer 1 ul 30 ul
10 uM Reverse primer 1 ul 30 ul
Taq polymerase 0,25 ul 7,5 ul
Nucease free hydrogen 41,75 ul 1252,5 ul
Total 50 ul 1500 ul


15 colonies have been randomly selected to be checked via PCR.

 

13

Assessing CooA Production

August 16

Testing gas production

August 16

Elise Grootscholten | Paul Reusink

Making a set up for the gas production testing and testing it with NEB10Bèta with the expected biobricks in it (K133071 + J23100) and a negative control.

Experiment 1: Gasproduction testing
15-08-18
Making a set up for the gas production testing and testing it with NEB10Bèta with the expected biobricks in it (K133071 + J23100) and a negative control.

Materials
-15 mL tubes
-Little glass tubes
-Cocktail prickers
-Culture medium
-Urea
-Chloramphenicol (40 mg/mL)


Methods
Stick tape on the little tube and cover the little tubes with tin foil. This can now enter the autoclave.
Make overnight cultures (20 mL)of the desired colonies in culture medium with 17,5 ul/ 20mL Chloramphenicol.

After a day, prepare the culture medium for the tests.
Put 1 g urea and 17,5 ul Chloramphenicol in 20 mL culture medium.

Pipette 10 mL of the overnight culture in the 15 mL tube en centrifuge 5000 rpm fot 5 minutes. Throw the supernatant away.
Fill the 15 mL tube and the little tube to the edge with culture medium. Work sterile.
Put a paper on the little tube and turn it around.
Then hold it above the 15 mL tube and pull the paper away and let the little tube fall into the 15 mL tube.
Try to do this so there's no gas in the little tube.
Now wait until gas is produced.

16-08-18
Making a set up for the gas production testing and testing it with NEB10Bèta with the expected biobricks in it (K133071 + J23100).
20 different colonies are tested.

Materials
Use the same materials as above.

Methods
Use the protocol above.
Make 700 mL culture medium with 35 g urea and 612,5 ul Chloramphenicol (40 mg/mL). This cannot be done sterile.
The culture medium can be used for the 20 different colonies with expected biobricks: K133071 + J23100.

 
Results
Gas was let into the little tubes so it was difficult to see whether there was gasproduction or not. Mainly in the negative control the tubes started with gas in it.

 

Figure 1: Little tube with cocktailpricker and 15 mL tube

Figure 2: Set up testing gas production

Figure 3. tube 1,2,3 and 4 starting amounts of air

Figure 4: After the weekend tube 1, 2, 3 and 4

Figure 5. tube 5,6,7 and 8 starting amounts of air

Figure 6: After the weekend tube 5, 6, 7 and 8

Figure 7. tube 9 and 10 strating amounts of air

Figure 8: After the weekend tube 9 and 10

Figure 9. tube 11,12,13,and 14 starting amounts of air

Figure 10: After the weekend tube 11, 12, 13 and 14

Figure 11. tube 15,16,17 and 18 starting amounts of air

Figure 12: After the weekend tube 15, 16, 17 and 18

Figure 13. tube 19 and 20 starting amounts of air

Figure 14: After the weekend tube 19 and 20

01

Testing gas production

August 16

Assessing Gas Production BioBricks in E.Coli

August 21

Elise Grootscholten | Randall de Waard

Minipreps are made of colonies 11 and 19 of biobrick combination K133071 + J23100. Results: 11. 270,57 ng/ul 19. 253,59 ng/ul.

Minipreps
21-08-18

Minipreps of colonies 11 and 19 (see Testing gasproduction experiment 1)

Materials
- Mini prep, Plasmid Purification Kit, Machery Michels 
- Eppendorf cups
- Variable Volume Pipets
- sterile pipet points
-Chloramphenicol (40 mg/mL)

Methods 
Miniprep protocol Plasmid Purification Kit, Machery Nagel
See experiment 3

Results

11. K133071 + J23100: 270,57 ng/ul
19. K133071 + J23100: 253,59 ng/ul

Figure 1: Results minipreps colonies 11 and 19

08

Assessing Gas Production BioBricks in E.Coli

August 21

Testing gas production

August 22

Elise Grootscholten | Paul Reusink

Making a set up for the gas production testing and testing it with NEB10Bèta with the expected biobricks in it (K133071 + I13453), (K173003 + J23100), (K173003 + I13453) and a negative control. Colonies 9 and 10 are chosen use for further tests because there was the biggest amount of gas production.

Experiment 2: Gasproduction testing
20-08-18

Making a set up for the gas production testing and testing it with NEB10Bèta with the expected biobricks in it (K133071 + I13453), (K173003 + J23100), (K173003 + I13453) and a negative control

Materials
See materials Experiment 1

Methods
See methods Experiment 1
850 mL culture medium is made with 21,5 g LB for 42 erlenmeyers (also for CooA production).
Overnight cultures are made of 10 colonies each of K173003 + I13453 and K173003 + J23100.

21-08-18

K173003 + J23100 are tested (see methods experiment 1)
Overnight cultures are made of  K133071 + I13453 and K173003 + I13453 are tested later.

Results
Colonies 9 and 10 are chosen use for further tests.

Figure 1. Starting amounts of air in tube 1,2,3 and 4.

Figure 2. Gas formation in tube 1,2,3 and 4. 30 minutes after adding Sodium pyruvate.

Figure 3. Starting amounts of air in tube 5,6,7 and 8.

Figure 4. Gas formation in tube 5,6,7 and 8. 30 minutes after adding Sodium pyruvate.

Figure 5. Starting amounts of air in tube 9 and 10.

Figure 6. Gas formation in tube 9 and 10. 30 minutes after adding Sodium pyruvate.

Figure 7: Gas formation in tube 1, 2, 3 and 4 after a dayr

Figure 8: Gas formation in tube 5, 6, 7 and 8 after a day

Figure 9: Gas formation in tube 9 and 10 after a day

02

Testing gas production

August 22

Testing gas production

August 22

Elise Grootscholten | Paul Reusink

Gasproduction testing for the biobricks (K173003 + I13453), (K133071 + I13453) and negative controls (B0015 and K133071 without urea and arabinose).
K173003 + I13453 is tested with sodiumpyruvate and arabinose for gasproduction and K133071 + I13453 is tested with urea and arabinose for gasproduction.
The negative control also produces a little bit gas.

Experiment 3: Gasproduction testing

Gasproduction testing for the biobricks K173003 + I13453, K133071 + I13453 and negative controls.
K173003 + I13453 is tested with sodiumpyruvate and arabinose for gasproduction and K133071 + I13453 is tested with urea and arabinose for gasproduction.

Materials
See experiment 1 and 2.

Methods
See experiment 1 and 2.
375 mL culture medium with CAM (already in it) + 18,75 g urea + 0,5 ul arabinose.
340 mL culture medium with CAM (already in it) + 51 mL natriumpyruvate (20 mM) + 0,5 ul arabinose.


Results and conclusion
Colonies 5 and 7 of the biobricks K173003 + I13453 (pyruvate)  and number 4 of the biobricks K133071 + I13453 (urea).
In both the negative controls there's been gas produced. Futher gasproduction test will be done.

Figure 1: Starting amounts in tube 1, 2, 3, 4 and 5 with Pyruvate and arabinose

Figure 2: Gasproduction in tube 1, 2, 3, 4 and 5 after a day with pyruvaat and arabinose

Figure 3: Starting amounts in tube 6, 7, 8, 9 and 10 with Pyruvate and arabinose

Figure 4: Gasproduction in tube 6, 7, 8, 9 and 10 after a day with pyruvate and arabinose

Figure 5: Starting amounts in tube 1, 2, 3, 4 and 5 with Urea and arabinose

Figure 6: Starting amounts in tube 6, 7, 8, 9 and 10 with Urea and arabinose

Figure 7: Gasproduction in tube 1, 2, 3, 4 and 5 after a day with urea and arabinose

Figure 8: Gasproduction in tube 6, 7, 8, 9 and 10 after a day with urea and arabinose

Figure 9: Starting amounts in the negative controls B0015 and K133071 + I13453 (without urea and arabinose)

Figure 10: Gasproduction in the negative controls B0015 and K133071 + I13453 after a day (without urea and arabinose)

03

Testing gas production

August 22

Testing gas production

August 24

Elise Grootscholten | Paul Reusink

Testing the gasproduction of the colonies 9 and 10 of biobricks (K173003 + J23100), colonies 5 and 7 of biobricks (K173003 + I13453) and colony 4 of biobricks (K133071 + I13453) with and without centrifuging the bacteria. There is also a negative control (J04450 pSB1K3) with Kanamycine. The negative contol started with a lot of gas inside the tube. We can not see wether there is produced more after a day or not. This have to be tested later.

Experiment 4: Gasproduction testing 
24-08-18

Testing the gasproduction with and without centrifuging the bacteria.

methods
See methods experiment 1
2 negative controls
colonies 9 and 10 of biobricks K173003 + J23100.
Colonies 5 and 7 of biobricks K173003 + I13453
Colony 4 of biobricks K133071 + I13453

Culture medium:
150 mL: Sodiumpyruvate (20 mM) 22,5 mL + Chloramphenicol (40 mg/mL) 131 ul
150 mL: Sodiumpyruvate (20mM) 22,5 mL +  Chloramphenicol (40 mg/mL) 131 ul + Arabinose (300 mg/mL) 0,5 ul
80 mL: Urea 4 g + Chloramphenicol (40 mg/mL) 70 ul + Arabinose (300 mg/mL) 0,5 ul
80 mL: Kanamycine (40 mg/mL) 70 ul

 

Figure 1: Starting amounts Nr. 9, 10 and 4 centrifuged

Figure 2. Gasproduction Nr 9. 10 and 4 after 4 hours centrifuged.

Figure 3: Starting amounts Nr. 5, 7 and negative control centrifuged

Figure 4. Gasproduction Nr 5,7 and negative control after 4 hours centrifuged.

Figure 5: Starting amounts Nr. 9, 10 and 4 not centrifuged

Figure 6. Gasproduction Nr 9. 10 and 4 after 4 hours not centrifuged.

Figure 7: Starting amounts Nr. 5, 7 and negative control not centrifuged

Figure 8. Gasproduction Nr. 5, 7 and negative control after 4 hours, not centrifuged.

04

Testing gas production

August 24

Urea and sodium pyruvate test for resistance E.coli

August 28

Randall de Waard | Elise Grootscholten

Testing different amounts of urea and sodiumpyruvate to know which concentrations the bacteria survive.

Experiment 2: Testing different concentrations of Urea and Sodium pyruvate
13-08-18


Same testing like experiment 1 but with different concentrations.

What How much Solution
Urea 1,2 g 1 mL purified water
Urea 1,6 g 2 mL purified water
Sodium pyruvate (10 mM) 10 mL None


Results
 
What How much Solution Result
Urea 1,2 g 1 mL purified water Little grow
Urea 1,6 g 2 mL purified water No grow
Sodium pyruvate (10 mM) 10 mL - Grow
Positive control - - Grow
Negative control - - No Grow
Table 2: Results experiment 2

Conclusion

For urea is 1 g used/20 mL and for sodiumpyruvate is 6 mL (10mM) or 3 mL (20 mM) used.
 
02

Urea and sodium pyruvate test for resistance E.coli

August 28

Assessing Gas Production BioBricks in E.Coli

August 30

Elise Grootscholten | Randall de Waard

A miniprep of the biobrick K352002 is made. The concentration is 69,77 ng/ul.

Experiment 9: Minipreps
28-08-18

Materials
- Mini prep, Plasmid Purification Kit, Machery Nagel
- Eppendorf cups
- Variable Volume Pipets
- sterile pipet points
-Chloramphenicol (40 mg/mL)

Methods
Miniprep protocol Plasmid Purification Kit, Machery Nagel
See experiment 3
Make minipreps of the biobricks K352002, K352003

Results
The minipreps can not be used, because washing buffer without ethanol is used. The miniprep of K352002 has been done again.

29-08-18

Methods
Make a miniprep of the biobrick K352002

Results
K352002: 69,77 ng/μL

30-08-18

Methods
See experiment 3
Make minipreps of the biobricks K133116 and K173013

Figure 1: Miniprep K352002

09

Assessing Gas Production BioBricks in E.Coli

August 30

Chemo competent cells

August 30

None

Competent cells of NEB10 Beta and BL21 DE3 were made. These cells were made competent so new biobricks and ligation mixes could be transformed into these cells. BL21 DE3 and NEB10 beta had 80 cups of 50 ul each. 1 cup is needed for 1 transformation later. The cells were competent enough to use in the project. 

Making chemocompetent cells: 

Materials: 
- E. coli NEB10Beta strain 
- E. coli Bl21 (DE 3) strain
- Buffer 1 

30 mM natriumacetaat, 100mM rubidium chloride,10 mM calciumchloride, 50mM mangaan chloride, 15 % glycerol; pH 5,8 met verdunde azijnzuur, filter steriliseren.

Bewaren bij kamertemperatuur.

- Buffer 2 

10 mM MOPS, 75 mM calcium chloride, 10 mM rubidium chloride, 15% glycerol; pH 6,6 met NaOH, filter steriliseren.

Bewaren bij kamertemperatuur 

Method: 
The day before making competent cells the strains were ented into 20ml LB medium (100ml erlenmeyer) and incubated for 6 hours by 37 degrees at 200 rpm. The culture was then transmitted to 20 ml LB medium (100 ml erlenmeyer) in a dilution of 1:50 (400 ul/ 20 ML) this was incubated overnight at 37 degrees Celsius at 200 rpm. 

In the morning the culture was diluted 1:100 in 200 ml LB medium (1L erlenmeyer). This was then incubated by 37 degrees and 225 rpm till a OD600 level of 0,5. 
To make sure the right OD was used, the culture was measured every 20/30 minutes. The results of the measurements can be found in part 2. 

After hitting the OD600 of 0,5. The cultures were transmitted to 250 ml centrifuge tubes and cooled on ice. The cells are supposed to stay on ice as much as possible after this. After cooling down to 4 degrees, the cultures were centrifuged at 4500 rpm for 10 min at 4 degrees. The supernatant were disposed, the pellet was resuspended in 66,4 ml ice cold Buffer 1(see materials). This was centrifuged  at 5000 rpm for 10 min at 4 degrees. 


BL21DE3  80 cups on place: Drawer 3, tower 2, drawer 3, place 2. 
NEB10Beta 79 cups on place: Drawer 3, tower 2, drawer 3, place 3. 

01

Chemo competent cells

August 30

Assessing Gas Production BioBricks in E.Coli

August 31

Elise Grootscholten | Randall de Waard

Digestions, gelelectrophoresis, dephosphorylations and ligations of different biobrick combinations. These biobrick combinations are 4 different promoters (J23100, I13453, K352002, K352003)  with 4 different gasproduction biobricks (k173003, K173013, K133071, K133116). J23100 is about 1 kb to long. The rest seems likely to be right digested. The ligations will be transformed in NEB10bèta and digested again as control.

Experiment 10: Digestion, defosforylation and ligation

Materials
-Cutsmart buffer (10x)
-Restriction enzymes XbaI, PstI-HF, EcoRI-HF, SpeI
-Nuclease free water
-DNA
-Phosphatase buffer (10x)
-Phosphatase (1 U/μL)
-Delutionbuffer (5x)

-T4 ligationbuffer (10x)
-T4 DNA ligase (1U/μL)

Methods
 

DNA (J23100 325,59 ng/μl) ~ 2 μg 6,1
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Restriction enzyme 2 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL

Table 1: Digestion scheme 1
 

DNA (J23100 325,59 ng/μl) ~ 2 μg 6,1
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL
Table 2: Digestion scheme 2
 
DNA (J23100 325,59 ng/μl) ~ 2 μg 6,1
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL
Table 3: Digestion scheme 3
 
DNA (K352003 180,65 ng/μl) ~ 2 μg 11,1 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 2 μL
Restriction enzyme 2 (1 U/μL) 2 μL
Nuclease free water Fill to 50 μL
Table 4: Digestion scheme 4
 
DNA (K352002 69,77 ng/μl) ~ 2 μg 28,7 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 2 μL
Restriction enzyme 2 (1 U/μL) 2 μL
Nuclease free water Fill to 50 μL
Table 5: Digestion scheme 5
 
DNA (K173003 217,06 ng/μL) ~ 2 μg 9,2 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 2 μL
Restriction enzyme 2 (1 U/μL) 2 μL
Nuclease free water Fill to 50 μL
Table 6: Digestion scheme 6
 
DNA (I13453 88,18 ng/μL) ~ 2 μg 22,7 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Restriction enzyme 2 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 7: Digestion scheme 7
 
DNA (K133071 186,79 ng/μL) ~ 2 μg 10,7 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Restriction enzyme 2 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 8: Digestion scheme 8
 
DNA (pSB1K3 163,93 ng/μL) ~ 2 μg  12,2 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 2 μL
Restriction enzyme 2 (1 U/μL) 2 μL
Nuclease free water Fill to 50 μL
Table 9: Digestion scheme 9
 
DNA (K173013 345,1 ng/μl) ~ 2 μg  5,8 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Restriction enzyme 2 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 10: Digestion scheme 10
DNA (K133116 194,1 ng/μl) ~ 2 μg 10,3 μL
Cutsmart (10x) 5 μL
Restriction enzyme 1 (1 U/μL) 1 μL
Restriction enzyme 2 (1 U/μL) 1 μL
Nuclease free water Fill to 50 μL
Table 11: Digestion scheme 11
 
Biobrick Restriction-enzyme 1 Restriction enzyme 2
K173003 XbaI PstI-HF
K173013 XbaI PstI-HF
K133071 XbaI PstI-HF
K133116 XbaI PstI-HF
J23100 EcoRI-HF SpeI-HF
I13453 EcoRI-HF SpeI-HF
K352002 EcoRI-HF SpeI-HF
K352003 EcoRI-HF SpeI-HF
pSB1K3 EcoRI-HF PstI-HF
Table 12: Digestion scheme used enzymes

Gelelectrophoresis
Number What Amount
1. Ladder 5 μL
2. J23100 EcoRI-HF+ SPeI-HF 5 μL
3. J23100 EcoRI-HF 5 μL
4. J23100 SpeI-HF 5 μL
5. pSB1K3 EcoRI-HF + PstI-HF 5 μL
6 K173013 XbaI + PstI-HF 5 μL
7. K173003 XbaI + PstI-HF 5 μL
8. K133071 XbaI + PstI-HF 5 μL
9. K133116 XbaI + PstI-HF 5 μL
10. K352002 EcoRI-HF + SpeI-HF 5 μL
11. K352003 EcoRI-HF + SpeI-HF 5 μL
12. I13453 EcoRI-HF + SpeI-HF 5 μL
13. Ladder  5 μL
14. Ladder 10 μL
Table 13: Gelelectrophoresis order of digestions

dephosphorylation

 

Volume Compound
12,5 μL Vector DNA (0,5 μg)
2 μL phosphatase buffer (10x)
4,5 μL milli Q
1 μL phosphatase (1 U/μL)
20 μL Total
Table 14: Pipetting scheme dephosphorylation
 
Volume Compound
37,5 μL Vector DNA (0,5 μg)
6 μL phosphatase buffer (10x)
13,5 μL milli Q
3μL phosphatase (1 U/μL)
60 μL Total
Table 15: Pipetting scheme dephosphorylation

Pre-Ligationmix:
Volume Compound
7,5 μL Insert DNA (2 μg/50 μL) 300 ng
7,5 μL Insert DNA (2 μg/50 μL) 300 ng
4 μL Backbone (0,5 μg/ 20 μL) 100 ng
5 μL Delutionbuffer (5x)
1 μL Milli Q
25 μL Total
Table 16: Pre-ligationmix pipette scheme

Ligationmix:
Volume Compound
10 μL Pre-ligationmix
2 μL 10x T4 ligationbuffer
1 μL T4 DNA ligase (1U/μL)
8 μL Milli Q
20 μL Total
Table 17: Ligationmix pipette scheme

Ligation started at 15.45 (31-08-18) at room temperature.
end ligation around 11.00 (01-09-18)
Store at -20 degrees Celcius.
 
10

Assessing Gas Production BioBricks in E.Coli

August 31

Assessing Gas Production BioBricks in E.Coli

September 3

Elise Grootscholten | Randall de Waard

Transformations of the biobrick combinations in NEB10bèta. The transformations are plated on kanamycine agar plates, because all the ligations were done in pSB1K3 (kanamycine resistence) backbone. Pink and with colonies appeared after incubation by 37 degrees Celcius for about 12h. The white colonies will be used for further experiments.

Experiment 11: transformations of the biobrick ombinations in NEB10bèta.
03-09-18

Materials
See materials experiment 5.
-Kanamycin (40 mg/mL)
-Ampicillin (50 mg/mL)


Methods
See methods experiment 5

Make 19 agar plates with kanamycine (Kana) (17,5 ul/20mL) and 1 agar plate with ampicillin (Amp) (40 ul/mL). Plate all the transformations on agar plates with Kana , because all the ligations are done in the pSB1K3 (Kana resistence) backbone. 
Devide one Amp plate and one Kana plate in about 30 parts. Because the biobrick combinations with biobrick K133116 have originally Amp and Kana resistence, the right backbone plasmid (with hopefully the right ligation) is only growing on the agar plate with Kana.


Results
There is grown n every transformation plates, so the transformations are done well.
There are pink and white colonies present. Only the white colonies will be used for further experiments.
 

11

Assessing Gas Production BioBricks in E.Coli

September 3

Soldering PCB [TC]

September 4

Tom van Dijk | Mike Vrieswijk
Soldered the first temperature controller with the help of a sodlering stencil and reflow oven. All components were soldered onto the first temperature controller.

01

Soldering PCB [TC]

September 4

Soldering PCB [TC]

September 4

Tom van Dijk | Mike Vrieswijk
Soldered second temperature controller with the help of a sodlering stencil and reflow oven. All components were soldered onto the first temperature controller.

02

Soldering PCB [TC]

September 4

Assessing Gas Production BioBricks in E.Coli

September 6

Elise Grootscholten | Randall de Waard

Plasmid DNA was isolated of 20 different colonies. The first time something went wrong. The second time we had good concentrations of isolated plasmid DNA. After this we will digest the DNA to see if the plasmids all have the required biobricks. The tube have a code from now on, see table 1.
 

Number Promotor Gene Backbone
A K352002 (CooF) K173013 pSB1K3
B K352002 (CooF) K173003 pSB1K3
C K352002 (CooF) K133071 pSB1K3
D K352003 (CooM) K133071 pSB1K3
E K352003 (CooM) K173003 pSB1K3
F K352003 (CooM) K173013 pSB1K3
H I13453 K173013 pSB1K3
I I13453 K173003 pSB1K3
J I13453 K133071 pSB1K3

Table 1: list of biobricks of abbreviations

Experiment 12: Overnight cultures and minipreps
05-09-18/06-09-18

Materials
See materials experiment 3

Methods
See methods experiment 3

Number Promotor Gene Backbone
A K352002 (CooF) K173013 pSB1K3
B K352002 (CooF) K173003 pSB1K3
C K352002 (CooF) K133071 pSB1K3
D K352003 (CooM) K133071 pSB1K3
E K352003 (CooM) K173003 pSB1K3
F K352003 (CooM) K173013 pSB1K3
G B0032 J23105 pSB1A3
H I13453 K173013 pSB1K3
I I13453 K173003 pSB1K3
J I13453 K133071 pSB1K3
Table 1: list of biobricks of abbreviations
 

Figure 1: Nanodrop results after DNA isolation. The biobrick combinations can be found in the methods (experiment 12)

12

Assessing Gas Production BioBricks in E.Coli

September 6

Assessing CooA Production

September 6

Dustin van der Meulen
After transforming the ligation product of J23105+B0032 and K352001+B0015 we had some growth of none red/pink colonies. Overnight cultures where made, and the DNA was prepped later on, checking them on gel.

Today a miniprep of pSB1K3 + J23100 + B0032 was performed. This gave concentrations of 51,3 and 89 ng/μL.

Of this, together with pSB1K3+ K352001 + B0015 and pSB1A3 a digest was performed:

 

Biobricks Enzyme 1 Enzyme 2
1K3+J23+B32 EcoRI SpeI
1K3+K35+B15 XbaI PstI
pSB1A3 EcoRI PstI

After two hours the digest was put on gel, though the rest was left for another two hours.

 
WhatsApp Image 2018-09-07 at 15.28.47.jpeg
 
 

CooA experiment 14

14

Assessing CooA Production

September 6

Assessing Gas Production BioBricks in E.Coli

September 10

Elise Grootscholten | Randall de Waard

The minipreps of experiment 12 are digested with the restriction enzymes: SmaI and ScaI. Only Sca and Sma are both incubated at 37 degrees Celcius. Sma has to be incubated at 25 degrees Celcius. This is done in experiment 15.

Experiment 12: Digestions of the minipreps
10-09-18

Materials
-SmaI (restriction-enzyme)
-ScaI (restriction-enzyme)
See materials experiment 10

Methods
See methods experiment 10
 

DNA (A1 396.7 ng/μl) ~ 2 μg 5.0 μL
Cutsmart (10x) 5 μL
Sca1-HF (2 U/μL) 1 μL
Sma1 (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
Table 1: Digestion scheme 1
 
DNA (B1 15.65 ng/μl) ~ 2 μg 127.8
Cutsmart (10x) 5 μL
Sca1-HF (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
Table 2: Digestion scheme 2
 
DNA (C1 186.6 ng/μl) ~ 2 μg 10.7 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 3: Digestion scheme 3
 
DNA (D1 79.0    ng/μl) ~ 2 μg 25.3 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 4: Digestion scheme 4
 
DNA (E1 97.7 ng/μl) ~ 2 μg 20.7 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 5: Digestion scheme 5
 
DNA (F1 106.7 ng/μl) ~ 2 μg 18.7 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 6: Digestion scheme 6
 
DNA (G1 328.9 ng/μl) ~ 2 μg 6.1 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 7: Digestion scheme 7
 
DNA (H1 121.0 ng/μl) ~ 2 μg 16.5 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 8: Digestion scheme 8
 
DNA (I1 1129.5 ng/μl) ~ 2 μg  1.7 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 9: Digestion scheme 9
 
DNA (J1 1465.7 ng/μl) ~ 2 μg  1.4 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 10: Digestion scheme 10
 
DNA (A2 310.4 ng/μl) ~ 2 μg 6.4 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 11: Digestion scheme 11
 
DNA (B2 364.9l) ~ 2 μg 5.5 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 12: Digestion scheme 12
 
DNA (C2 228.3 ng/μl) ~ 2 μg 8.8 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 13: Digestion scheme 13
 
DNA (D2 701.7 ng/μl) ~ 2 μg 2.9 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 14: Digestion scheme 14
 
DNA (E2 119.8 ng/μl) ~ 2 μg 16.7 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 15: Digestion scheme 15
 
DNA (F2 493.3 ng/μl) ~ 2 μg 4.1 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 16: Digestion scheme 16
 
DNA (G2 211.7 ng/μl) ~ 2 μg 9.5 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 17: Digestion scheme 17
DNA (H2 482.2 ng/μl) ~ 2 μg 4.2 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 18: Digestion scheme 18
 
DNA (I2 1364.7 ng/μl) ~ 2 μg 1.5 μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
 Table 19: Digestion scheme 19
 
DNA (J2 1665.5 ng/μl) ~ 2 μg1.2  μL
Cutsmart (10x) 5 μL
Sca1-HF 1 (2 U/μL) 1 μL
Sma1  (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL
Table 20: Digestion scheme 20



Results
The results of the digestions can be found in figure 1 and 2 (Images experiment 13)

Ladder, A1, B1, C1, D1, E1, F1, G1, H1, I1, (J1), Ladder, A2, B2, C2, D2, E2, F2, G2, Ladder
Ladder, H2, I2, J2, Ladder, K3, J1, ladder
Something went wrong with the first (J1), so look at the second one.

Conclusion/discussion
Restriction-enzyme Sma was incubated at 37 degrees Celcius (same as Sca). This must be 25 degrees Celcius.
Therefore, Sma was added again and was incubated at 25 degrees Celcius. See results experiment 16.

Figure 1: Digestions with ScaI and SmaI

Figure 2: Digestions with ScaI and SmaI

13

Assessing Gas Production BioBricks in E.Coli

September 10

Soldering PCB [TC]

September 11

Tom van Dijk | Mike Vrieswijk
Testing first temperaturecontroller. Reworking surton components due to shorts on the PCB. Rework succesfull.

03

Soldering PCB [TC]

September 11

Soldering PCB [TC]

September 11

Tom van Dijk | Mike Vrieswijk
Testing second temperaturecontroller. Reworking surton components due to shorts on the PCB. Rework failed.

04

Soldering PCB [TC]

September 11

Assessing Gas Production BioBricks in E.Coli

September 13

Elise Grootscholten | Randall de Waard

Minipreps of the biobrick combinations: K352002+K133116+pSB1K3 (1) and K352003+K133116+pSB1K3 (2) are done. The DNA concentration is for 1:189,3 ng/μl and for 2: 355,3 ng/μl.

Experiment 14: Minipreps
13-09-18

Minipreps of the biobrick combinations: K352002+K133116+pSB1K3 and K352003+K133116+pSB1K3

Materials
- Mini prep, Plasmid Purification Kit, Machery Nagel
- Eppendorf cups
- Variable Volume Pipets
- sterile pipet points
-Chloramphenicol (40 mg/mL)

Methods
Machery Nagel Plasmid Purification
See experiment 3

Results
K352002+K133116+pSB1K3: 189,3 ng/μl
K352003+K133116+pSB1K3: 355,3 ng/μl

Figure 1: Minipreps 13-09-18

14

Assessing Gas Production BioBricks in E.Coli

September 13

Assessing Gas Production BioBricks in E.Coli

September 13

Elise Grootscholten | Randall de Waard

Digestions have been done of the biobrickcombinations K352002 +K133116 + pSB1K3 and K352003 + K133116 + pSB1K3 with restriction-enzymes ScaI and SmaI

Experiment 14: Digestions
13-09-18

Digestions of the biobrick combinations: K352002+K133116+pSB1K3 and K352003+K133116+pSB1K3

Materials
-SmaI (restriction-enzyme)
-ScaI (restricion-enzyme)
See experiment..


Methods
1 μL SmaI restriction-enzyme has been added to the digetions from experiment 13, except for pSB1K3.
After an our at room temperature the digestions were put in -20 degrees Celcius.

 
DNA (K352002+K133116+pSB1K3 189,3 ng/μl) ~ 2 μg 32,4 μL
Cutsmart (10x) 5 μL
Sca1-HF (2 U/μL) 1 μL
Sma1 (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL

Table 1: Digestion scheme 1

DNA (K352003+K133116_pSB1K3 355,3 ng/μl) ~ 2 μg 5,6 μL
Cutsmart (10x) 5 μL
Sca1-HF (2 U/μL) 1 μL
Sma1 (2 U/μL) 1 μL
Nuclease free water Fill to 50 μL

Table 2: Digestion scheme 2

 

 

15

Assessing Gas Production BioBricks in E.Coli

September 13

Assessing Gas Production BioBricks in E.Coli

September 14

Elise Grootscholten | Randall de Waard

All the digestions of experiment 13 have digested again with SmaI for an our at 25 degrees Celcius. Because earlier the digestions have been put immediately bt 37 degrees Celcius. It looks like H1, J1 and D2 include the right biobricks. 

Experiment 16: Gelelectroforesis
14-09-18

Materials
-Loading dye (6x)
-2 Log DNA ladder
-Digested DNA
-1% arabinose gel (1 g/100 mL TAE buffer)

Methods
Put 1 gram of arabinose with 100 mL TAE buffer.
Heat it in the microwave until the solution becomes clear.
Add 100 μL gelred when the solution is hand warm.
Pour the solution into the gel holder which you taped before.
Wait until the gel is solid then you can pipette your dna with Loading dye into the wells.

L: 3 μL
DNA: 2 μL loading dye + 5 μL DNA, 5 μL on gel.

Gel 1
L, A1, B1, C1, D1, E1, F1, G1, G1 with only smaI, H1, I1, J1, A2, B2, C2, D2, E2, F2, G2, L

Gel 2
L, H2, I2, J2, L, K352002 + K133116 + pSB1K3,  K352003 + K133116 + pSB1K3

Figure 1: Gel 1 (14-09-18), digested with ScaI and SmaI.

Figure 2: Gel 2, H2, I2, J2, L, K352002+K133116+pSB1K3, K352003+K133116+pSB1K3

16

Assessing Gas Production BioBricks in E.Coli

September 14

Testing gas production

September 17

Elise Grootscholten | Paul Reusink

Different strains of E.coli are tested as negative control. Those strains were: NEB10Bèta, BL21 (DE3), BL21, HB101, DH5alpha and JM109. NEB10Bèta produced the most gas. BL21, HB101 and JM101 produced none/almost none gas. Because BL21 isn't a K12 strain we can not use that one. That is why we will test further gasproduction in HB101 and JM109. And maybe those will be our final E.coli strains.

Experiment 5: Testing different strains of E.coli

Materials
All the following types of bacteria are E.coli:
-glycerolstock BL21
-glycerolstock HB101
-glycerolstock DH5alpha
-glycerolstock JM109
-agarplate with NEB10beta
-agarplat with BL21 (DE3) 

Methods
See experiment 1
Make overnight cultures of the following E.coli tribes: NEB10Bèta, BL21 (DE3), BL21, HB101, DH5alpha, JM109.
All the bacteria are de deluted in normal LB agar, LB agar with urea and LB agar with pyruvate. 


Results
 

Bacteria LB LB Urea Urea Pyruvate Pyruvate
NEB10beta + +++ +++ +++ + ++/+++
BL21 (DE3)  - ++ - - - -
BL21 - - - - - -
DH5a +++ +/- - - +/- +
HB101 - - - - + -
JM109 - - - - - -
05

Testing gas production

September 17

Designing peristaltic pump [PP]

September 18

Tom van Dijk | Mike Vrieswijk
Made a 3d-model of the parts for the peristaltic pump. Parts include: multiple gears, bearings, and casing.

01

Designing peristaltic pump [PP]

September 18

3d-printing peristaltic pump [PP]

September 18

Tom van Dijk | Mike Vrieswijk
3d-printed all parts of the peristaltic pump model.

02

3d-printing peristaltic pump [PP]

September 18

Animating peristaltic pump [PP]

September 18

Tom van Dijk | Mike Vrieswijk
An animation was made of the first peristaltic pump.

03

Animating peristaltic pump [PP]

September 18

Assessing CooA Production

September 20

Dustin van der Meulen

This week we peformed another cycle of cloning, starting with digesting J23105+B0032 with EcoRI and SpeI and K352001+B0015 with XbaI and PstI, checking these on gel (which seemed to be correct) and ligating them together on the pSB1A3 version of BBa_J04450. This was then grown overnight and prepped the following day.

Today a dephos and ligation has been performd using the rAPId dephos and ligation kit and protocol.
For the dehpos a total of 17 μL of pSB1A3 wasused, for an end concentration of 17 ng/μL
For the ligationthe endvolume was trippled as to have enough DNA.
For both inserts (K352001 & B0015 | J23100 + B0032) 7.5 μL was used (150 ng of DNA foreach insert)
For the veccor (pSB1A3) 3 μL was used (51 ng of DNA)
The ligation will be left overnigh (started at 15:30).


 

15

Assessing CooA Production

September 20

Assessing Gas Production BioBricks in E.Coli

September 24

Elise Grootscholten | Randall de Waard
32 minipreps have been performed. A lot of them have a low concentration nucleic acid and/or have a high 260/280 and 260/230 rate. Therefore those will be done again.

Experiment 17: Minipreps
20-09-18
Overnight cultures (35)
21-09-18
Minipreps (32)

Materials 
See experiment 3
-Kanamycin (40 mg/mL)

Methods
See experiment 3
Make 32 minipreps:
A: 6
B: 6
C:6
D:1
E:6
F:6
H:1
I: 1
J:2

Results/conclusion
There was no grow in B6, C2 and C5.
A lot of them have a low concentration nucleic acid and/or have a high 260/280 and 260/230 rate. Therefore those will be done again.
Thise are: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5

Figure 1: Minipreps, a total of (32) because number 5, 7 and 12 are wrong.

17

Assessing Gas Production BioBricks in E.Coli

September 24

Integration of silicon hose and peristaltic pump [PP]

September 25

Tom van Dijk | Mike Vrieswijk
Due to ordering issues a different silicon hose was implemented in the design. The new silicon tube was integrated into the 3d-printed peristaltic pump. A test was done with the new silicon tube in place. The test cocluded that the tube was not compatible with this peristaltic pump design. A new design must be made.

01

Integration of silicon hose and peristaltic pump [PP]

September 25

Designing peristaltic pump v2 [PP]

September 25

Tom van Dijk | Mike Vrieswijk
Peristaltic pump was redesigned and the model has been updated.

02

Designing peristaltic pump v2 [PP]

September 25

3d-printing peristaltic pump v2 [PP]

September 25

Tom van Dijk | Mike Vrieswijk
3d-printed all parts of the peristaltic pump model v2. All parts have been put together succefully.

03

3d-printing peristaltic pump v2 [PP]

September 25

Testing peristaltic pump v2 [PP]

September 25

Tom van Dijk | Mike Vrieswijk
The silicon tube was intergrated with the 3e-printed peristaltic pump v2 and tested. The test concluded that the motor had insufficient force. A new design must be made.

04

Testing peristaltic pump v2 [PP]

September 25

Designing peristaltic pump v3 [PP]

September 26

Tom van Dijk | Mike Vrieswijk
An editional planetary gearset was added to the first planetary gearset of the peristaltic pump v2 model to increase the power output by 4x.

01

Designing peristaltic pump v3 [PP]

September 26

3d-printing peristaltic pump v3 [PP]

September 26

Tom van Dijk | Mike Vrieswijk
3d-printed all parts of the peristaltic pump model v3. All parts have been put together succefully.

02

3d-printing peristaltic pump v3 [PP]

September 26

Testing peristaltic pump v3 [PP]

September 26

Tom van Dijk | Mike Vrieswijk
The newly pot together peristalticpump model v3 was tested. The test concluded a flaw in the design of the peristaltic pump v3. Failure: the silicon tube was pulled through the pump. A new design must be made.

03

Testing peristaltic pump v3 [PP]

September 26

Assessing CooA Production

September 26

Dustin van der Meulen
Digesting the CooA combinations with NotI we could check if the requiered biobricks where present in our cells. Out of 10 checked colonies, 3 appeared to be the right size. Because of an error the on plate location of these colonies was lost however, so another check will have to be performed.

Pipet 8 digestions
Nr. 1, 2, 4, 5, 7, 8, 9, and 10 of CooA. Use 5 μL of digest with 3 μL of loading dye.

Gel:
Ladder 5 ul 1, 2, 4, 5, 7, 8, 9, 10, pSB1K3 (Sca), G1 (Sma), Ladder 5 ul, ladder 3 ul
The CooA digestions are 5 ul
 

Gel CooA.jpg

 
 

CooA 18-09

16

Assessing CooA Production

September 26

Designing Test tube shaker [TTS]

September 27

Tom van Dijk | Mike Vrieswijk
A test tube shaker was designed to fit on top of a NEMU17 stepper motor. A spot for end switch was put in place to calibrate the test tube shaker. A cap for the test tube was designed.

01

Designing Test tube shaker [TTS]

September 27

3d-printing Test tube shaker [TTS]

September 27

Tom van Dijk | Mike Vrieswijk
The test tube shaker was 3d-printed and assembled. Nine caps were printed.

02

3d-printing Test tube shaker [TTS]

September 27

Testing Test tube shaker [TTS]

September 27

Tom van Dijk | Mike Vrieswijk
The newly assembled testtube shaker was tested for sufficient and controlled shakage. The test was deemed a succes. The calibration for the test tube shaker potition was tested and deemed in working order. The caps were tested for sufficient air permeability. The test concluded suficient air permeability.

03

Testing Test tube shaker [TTS]

September 27

Assessing Gas Production BioBricks in E.Coli

September 27

Elise Grootscholten | Randall de Waard

Making overnight cultures of the numbers: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5 with Kanamycin. These biobrick combinations can be found in experiment 12. Also the first digestions with XbaI, Eco0109I, HindIII and SspI-HF have been done.

Experiment 19: Making overnight cultures + first digestions
27-09-18

Materials
-Kanamycine 
-XbaI
-SspI-HF
-Eco019I
-HindIII

Methods
Make new overnight cultures of the numbers: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5 with Kanamycin. 

 
19

Assessing Gas Production BioBricks in E.Coli

September 27

Assessing Gas Production BioBricks in E.Coli

September 27

Elise Grootscholten | Randall de Waard

Minipreps have been made of the numbers: A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5.

Experiment 20: Minipreps of gasproduction biobricks
27-09-18

Make minipreps of A1, A4, A5, A6, B1, B5, B6 C1, C2, C3, C4, C5, C6, E1, E2, E5, E6, F4, F5.
See experiment 3.

Also make master mixes for the control digestions and code the tubes.
 

20

Assessing Gas Production BioBricks in E.Coli

September 27

Designing peristaltic pump v4 [PP]

September 28

Tom van Dijk | Mike Vrieswijk
The innershape of the tube duct was rounded to increase friction on the tube. The model of the peristaltic pump v3 was updated.

01

Designing peristaltic pump v4 [PP]

September 28

3d-printing peristaltic pump v4 [PP]

September 28

Tom van Dijk | Mike Vrieswijk
The new parts for the tubeduct were 3d-printed and put together with the peritstaltic pump.

02

3d-printing peristaltic pump v4 [PP]

September 28

Testing peristaltic pump v4 [PP]

September 28

Tom van Dijk | Mike Vrieswijk
The new peristaltic pump v4 was tested. The test concluded a large improvement over the previous generation, but was deemed unreliable due tothe tube occasional being pulled through the pump. A new design must be made.

03

Testing peristaltic pump v4 [PP]

September 28

Designing peristaltic pump v4.1 [PP]

September 28

Tom van Dijk | Mike Vrieswijk
A clamp was designed to hold the tube into the peristaltic pump from both ends. the peristaltic pump model v4 was updated.

04

Designing peristaltic pump v4.1 [PP]

September 28

3d-printing peristaltic pump v4.1 [PP]

September 28

Tom van Dijk | Mike Vrieswijk
The new clamp was 3d-printed, put together and added to the epristaltic pump.

05

3d-printing peristaltic pump v4.1 [PP]

September 28

Testing peristaltic pump v4.1 [PP]

September 28

Tom van Dijk | Mike Vrieswijk
The new peristaltic pump v4.1 was tested. The test conclude no noticable difference to the behaviour of the tube. The tube was still occsionally pulled through the pump. A new design must be made.

06

Testing peristaltic pump v4.1 [PP]

September 28

Designing peristaltic pump v5 [PP]

September 30

Tom van Dijk | Mike Vrieswijk
The tube clamp was attached to the top piece of the model's casing. The model of the persitaltic pump v4.1 was updated.

01

Designing peristaltic pump v5 [PP]

September 30

3d-printing peristaltic pump v5 [PP]

September 30

Tom van Dijk | Mike Vrieswijk
The new integrated tube clamp was 3d-printed and added to the peristaltic pump.

02

3d-printing peristaltic pump v5 [PP]

September 30

Testing peristaltic pump v5 [PP]

September 30

Tom van Dijk | Mike Vrieswijk
The new peristaltic pump v5 was tested. The new v5 was deemed a succes. No previous bad behaviour was observed.

03

Testing peristaltic pump v5 [PP]

September 30

Assessing CooA Production

October 1

Dustin van der Meulen
Today a colony PCR was performed of multiple CooA ligation products of J23100+B0032+K352001+B0015 on pSB1A3. This specific colony was later PCRed again with the gasproduction biobricks in preperation for sequencing. Only one of the colonies seemed to be the right length, though te lenght of the complete insert and BBa_J04450 is quite similair, so sequencing later this week will provide the final answer.

 

CooA 4-10

17

Assessing CooA Production

October 1

Assessing Gas Production BioBricks in E.Coli

October 4

Elise Grootscholten | Randall de Waard

From 27-09 until 04-10 more than 200 digestions and 17 gelelectrophoresis have been performed. This was done as a control for our composite parts. We digested with XbaI, Eco01019 and SspI-HF/HindIII. Every restriction-enzyme has an unique place to cut in our constructs. With this we could test if the promoter, backbone and gas production gene was present. However, the digestions didn't show us very good results. Only I1 and J1 looked good. After this we started with PCR.

Experiment 21: Digestions and gelelectrophoresis
01-10-18

Digestions of the gasproduction biobrick combinations (D, H, I and J) and a start with the gelelectrophoresis.

Methods
Digestions of D2, H1, I2 and J2 of experiment 12.
J2 will from now on be J3.

NEB 2.1 (10x) 4 μL
Eco0109I (20 U/μL) 1 μL
Nuclease free water 35 μL

Table 1: Master mix digestion 1
 

NEB 2.1 (10x) 4 μL
Eco0109I (20 U/μL) 1 μL
XbaI (20 U/μL) 1 μL
Nuclease free water 34 μL
 
Table 2: Master mix digestion 2
 
NEB 2.1 (10x) 2 μL
Eco0109I (20 U/μL) 1 μL
XbaI (20 U/μL) 1 μL
HindIII (20 U/μL) 1 μL
Nuclease free water 15 μL
Table 3: Master mix digestion 3
 
NEB 2.1 (10x) 2 μL
Eco0109I (20 U/μL) 1 μL
XbaI (20 U/μL) 1 μL
SspI-HF 1 μL
Nuclease free water 15  μL
Table 4: Master mix digestion 4
 
NEB 2.1 (10x) 2 μL
HindIII (20 U/μL) 1 μL
XbaI (20 U/μL) 1 μL
Nuclease free water 16 μL
 
Table 5: Master mix digestion 5
 
NEB 2.1 (10x) 2 μL
SspI-HF (20 U/μL) 1 μL
XbaI (20 U/μL) 1 μL
Nuclease free water 16 μL
 
Table 6: Master mix digestion 6
 
NEB 2.1 (10x) 4 μL
XbaI (20 U/μL) 1 μL
Nuclease free water 35 μL
 
Table 7: Master mix digestion 7
 
NEB 2.1 (10x) 2 μL
HindIII (20 U/μL) 1 μL
Nuclease free water 17 μL
 
Table 8: Master mix digestion 8
 
NEB 2.1 (10x) 2 μL
SspI-HF (20 U/μL) 1 μL
Nuclease free water 17 μL
 
Table 9: Master mix digestion 9
 
NEB 2.1 (10x) 2 μL
Eco0109I (20 U/μL) 1 μL
HindIII (20 U/μL) 1 μL
Nuclease free water 16 μL
 
Table 10: Master mix digestion 10
 
NEB 2.1 (10x) 2 μL
Eco0109I (20 U/μL) 1 μL
SspI-HF (20 U/μL) 1 μL
Nuclease free water 16 μL
 
Table 11: Master mix digestion 11

Pipette 9 μL of the mastermix with 2 μL of H1/ 1 μL of D2/ 1 μL of I2/ 1 μL of J23

D2 and J3 are digested with mastermix: 1, 2, 3, 5, 7, 8 and 10.
H1 and I2 are digested with mastermix: 1, 2, 4, 6, 7, 9, and 11.

L= 2 log DNA ladder
 

Gelelectrophoresis
01-10-18

Gel 1
L, B6 Eco01090I, B6 SspI-HF, B6 XbaI, B6 Eco0109I + SspI-HF, B6 Eco0109I + XbaI, B6 SspI-HF + XbaI, B6 Eco0109I + SspI-HF + XbaI, L, B2 SspI-HF, B2 XbaI, B2 Eco0109I + SspI-HF, B2 Eco0109I + XbaI, B2 SspI-HF + XbaI, B2 Eco0109I + SspI-HF + XbaI, L, H1 SspI-HF, H1 XbaI, H1Eco0109I + SspI-HF, H1 Eco0109I + XbaI

Gel 2
L, H1 SspI-HF + XbaI, H1 Eco0109I + SspI-HF + XbaI, I1 Eco0109I + SspI-HF, I1 Eco0109I + XbaI, I1 SspI-HF + XbaI, I1 Eco0109I + SspI-HF + XbaI, L, I1 SspI-HF, I1 XbaI, B3 SspI-HF, B3 XbaI, B3 Eco0109I + SspI-HF, B3 Eco0109I + XbaI, B3 SspI-HF + XbaI, B4 SspI-HF, B4 XbaI, B4 Eco0109I + SspI-HF, B4 Eco0109I + XbaI, B4 Eco0109I + SspI-HF + XbaI

Gel 3
L, F6 SspI-HF, F6 XbaI, F6 Eco0109I + SspI-HF, F6 Eco0109I + XbaI, F6 SspI-HF + XbaI, F6 Eco0109I + SspI-HF + XbaI, L, F5 SspI-HF, F5 XbaI, F5 Eco0109I + SspI-HF, F5 Eco0109I + XbaI, F5 SspI-HF + XbaI, F5 Eco0109I + SspI-HF + XbaI, L F4 XbaI, F4 Eco0109I + SspI-HF, F4 Eco0109I + XbaI, F4 sspI-HF + XbaI, L

Gel 4
L, F4 Eco0109 + XbaI + SspI-HF, F4 Eco0109, F3 XbaI, F3 Eco0109 + SspI-HF, F3 Eco0109 + XbaI, F3 SspI-HF + XbaI, F3 Eco0109 + XbaI + SspI-HF, F2 SspI-HF, F2 XbaI, L, F2 Eco0109 + SspI-HF, F2 Eco0109 + XbaI, F2 SspI-HF + XbaI, F2 Eco01090 + XbaI + SspI-HF, F1 XbaI, F1 Eco0109 + SspI-HF, F1 Eco0109 + XbaI, F1 SspI-HF + XbaI

Gel 5
L, F1 Eco0109 + XbaI + SspI-HF, E1 Eco0109, E1 SspI-HF, E1 XbaI, E1 Eco0109 + SspI-HF, E1 Eco0109 + XbaI, E1 SspI-HF + XbaI, E1 Eco0109 + XbaI + SspI-HF, L, E2 Eco0109, E2 SspI-HF, E1 XbaI, E2 Eco0109 + SspI-HF, E2 Eco0109 + XbaI, E2 SspI-HF + XbaI, E2 Eco0109 + XbaI + SspI-HF, L

Gel 6
L, E3 SspI-HF, E3 XbaI, E3 Eco0109 + SspI-HF, E3 Eco0109 + XbaI, E3 SspI-HF + XbaI, E3 Eco0109 + XbaI + SspI-HF, L, E6 Eco0109, E6 SspI-HF, E6 XbaI, E6 Eco0109 + SspI-HF, E6 Eco0109 + XbaI, E6 SspI-HF + XbaI, E6 Eco0109 + XbaI + SspI-HF, L

02-10-18
Gelelectrophoresis

Gel 1
L,- , A1 Eco0109 , A1 SspI-HF, A1 XbaI, A1 Eco0109 + SspI-HF, A1 Eco01019 + XbaI, A1 SspI-HF + XbaI, A1 Eco0109 + XbaI + SspI-HF, -, -, A2 Eco0109 , A2 SspI-HF, A2 XbaI, A2 Eco0109 + SspI-HF, A2 Eco01019 + XbaI, A2 SspI-HF + XbaI, A2 Eco0109 + XbaI + SspI-HF, -, L

Gel 2
L,- , A3 Eco0109 , A3 SspI-HF, A3 XbaI, A3 Eco0109 + SspI-HF, A3 Eco01019 + XbaI, A3 SspI-HF + XbaI, A3 Eco0109 + XbaI + SspI-HF, -, -, A4 Eco0109 , A4 SspI-HF, A4 XbaI, A4 Eco0109 + SspI-HF, A4 Eco01019 + XbaI, A4 SspI-HF + XbaI, A4 Eco0109 + XbaI + SspI-HF, -, L


03-10-18
Gelelectrophoresis

Gel 1
L, D2 Eco0109, D2 Eco0109 + XbaI, D2 Eco0109 +XbaI + HindIII, D2 HindII + XbaI, D2 XbaI, D2 HindIII, D2 Eco0109 + HindIII, D not digested, L, J3 Eco0109, J3 Eco0109 + XbaI, J3 Eco0109 + XbaI + HindIII, J3 XbaI, J3 HindIII, J3 Eco0109 + HindIII, J3.10, J not digested, L.

Gel 2
L, H2 Eco0109, H2 Eco0109 + XbaI, H2 Eco0109 + XbaI + SspI-HF, H2 SspI-HF + XbaI, H2 XbaI, H2 SspI-HF, H2 Eco0109 + SspI-HF, H2 not digested, L, I2 Eco0109, I2 Eco0109 + XbaI, I2 Eco0109 + XbaI + SspI-HF, I2 SspI-HF + XbaI, I2 XbaI, I2 SspI-HF, I2 Eco0109 +SspI-HF, I2 not digested, L

Gel 3
L, C1 Eco0109, C1 HindIII, C1 XbaI, C1 Eco0109 + XbaI, C1 Eco0109 + HindIII, C1 HindIII + XbaI, C1 Eco0109 + XbaI + HindIII, C1 not digested, L, C2 Eco0109, C2 HindIII, C2 XbaI, C2 Eco0109 + XbaI, C2 Eco0109 + HindIII, C2 HindIII + XbaI, C2 Eco0109 + XbaI + HindIII, C2 not dgested, L

Gel 4
L, C3 Eco0109, C3 HindIII, C3 XbaI, C3 Eco0109 + XbaI, C3 Eco0109 + HindIII, C3 HindIII + XbaI, C3 Eco0109 + XbaI + HindIII, C3 not digested, L, C4 Eco0109, C4 HindIII, C4 XbaI, C4 Eco0109 + XbaI, C4 Eco0109 + HindIII, C4 HindIII + XbaI, C4 Eco0109 + XbaI + HindIII, C4 not dgested, L

Gel 5
L, C6 Eco0109, C6 HindIII, C6 XbaI, C6 Eco0109 + XbaI, C6 Eco0109 + HindIII, C6 HindIII + XbaI, C6 Eco0109 + SspI-HF + XbaI, C6 not digested, L, B5 Eco0109, B5 SspI-HF, B5 XbaI, B5 XbaI (other tube), B5 Eco0109 + XbaI, B5 SspI-HF + Eco0109, B5 SspI-HF + XbaI, B5 Eco0109 + SspI-HF + XbaI, L

04-10-18
Gelelectrophoresis

Gel 1
L, J2 Eco0109, J2 HindIII, J2 XbaI, J2 Eco0109 + XbaI, J2 Eco0109 + HindIII, J2 HindIII + XbaI, J2 Eco0109 + HindIII + XbaI, J2 not digested, L, J1 Eco0109, J1 HindIII, J1 XbaI, J1 Eco0109 + XbaI, J1 Eco0109 + HindIII, J1 HindIII + XbaI, J1 Eco0109 + HindIII + XbaI, J1 not digested, L

Gel 2
L, B2 Eco0109, B2 HindIII, B3 Eco0109, B3 HindIII, B3 Eco0109 + SspI-HF + XbaI, B4 Eco0109, B4 HindIII, B4 Eco0109 + SspI-HF + XbaI, not digested B2, L, D1 Eco0109, D1 HindIII, D1 XbaI, D1 Eco0109 + XbaI, D1 Eco0109 + HindIII, D1 HindIII + XbaI, D1 Eco0109 + HindIII + XbaI, not digested D1, L

Gel 3
L, F1 Eco0109, F1 HindIII, F2 Eco0109, F3 Eco0109, F3 HindIII, F6 Eco0109, F6 HindIII not digested F6, L, A2 HindIII, A3 HindIII, not digested A1, E3 Eco0109, E3 HindIII, not digested E3, L

Gel 4
L, H1 Eco0109, H1 HindIII, H1 not digested, I1 Eco0109, I1 HindIII, L

Figure 1: Gel 1 01-10-18

Figure 2: Gel 2 01-10-18

Figure 3: Gel 3 01-10-18

Figure 4: Gel 4 01-10-18

iFigure 5: Gel 5 01-10-18

iFigure 6: Gel 6 01-10-18

Figure 7: Gel 1 02-10-18

Figure 8: Gel 2 02-10-18

Figure 9: Gel 1 03-10-18

Figure 10: Gel 2 03-10-18

Figure 11: Gel 3 03-10-18

Figure 12: Gel 4 03-10-18

Figure 13: Gel 5 03-10-18

Figure 14: Gel 1 04-10-18

Figure 15: Gel 2 04-10-18

Figure 16: Gel 3 04-10-18

Figure 17: Gel 4 04-10-18

21

Assessing Gas Production BioBricks in E.Coli

October 4

Preparing DNA for Submission

October 5

Dustin Vermeulen | Elise Grootscholten | Randall de Waard

Today we made colony PCRs of the biobricks we suspected that where correct of our gasproduction, and of colony 8 of CooA. These where then send to the Erasmus Medical Centre, who sequenced our parts for us.

Colony PCR
Materials: For our PCR (Both plasmid and colony) reactions we have used the standard neb protocols for a 25 μl reaction , with the following concentrations:

10X Standard Taq Reaction Buffer        2.5 μl
10 mM dNTPs                                        0.5 μl
10 μM forward Primer  (VF2)                 0.5 μl
10 μM reverse Primers  (VR)                 0.5 μl
Taq DNA Polymerase                             0.125 ul
Nuclease-free water                               to 25 μl

The primers VF2 and VR got delivered in powder form. So it was dissolved in Nuclease-free water till a concentration of 100 μm.

Methods: A master mix is made with the concentrations shown above. In case of a plasmid pcr, a volume of plasmid is added before the reaction is filled with Nuclease-free water. With the colony PCR we used a toothpick to touch on the desired colony. Before briefly submerging and shaking the tip of the toothpick in the PCR Eppendorfcup filled with the mastermix. Before pricking it into a clean Agar plate a couple of times to grow more colonies in case the PCR shows us the desired products

For our PCR reactions we use the Bio-Rad T100 thermal cycler. Afterwards the products of the PCR reaction were ran on a gel electrophoresis.

PCR Purification
8 μL PCR product + 2 μL ExoSAP-IT
37°C 20 minutes
80°C 15 minutes

 

Sequencing Reaction

4,0 μL PCR product
0,15 μL forward or reverse primer
0,5 μL BigDye V3.1 (Applied Biosystems)
1,75 μL Sequencing buffer (5x)
3,6 μL Nuclease free water
10 μL Total


96°C 1 minute
96°C 10 seconds
55°C 5 seconds
60°C 2 minutes
run for 35 cycli
10°C end

Sequencing reaction purification
Following protocol of CleanSeq Agencourt

Sequencing
ABI 3730 XL, 50 cm capillair

01

Preparing DNA for Submission

October 5

Preparing DNA for Submission

October 7

Dustin Vermeulen | Elise Grootscholten | Randall de Waard

Today we digested our succesfully sequenced biobricks (B1 / B6 (same biobrick), D1, I1 and J2), using EcoRI and PstI to get them into a pSB1C3 backbone. This was then transormed and plated. And the end of the day we made 6 overnight cultures of the barely visible colonies.

Materials
-Restriction enzymes EcoRI-HF and PstI

Methods
Digestion

Mastermix

Cutsmart (10x) 55 μL
EcoRI-HF (20 U/μL) 1 μL
PstI (20 U/μL) 1 μL
Nuclease free water 382,7 μL


50 μL mastermix - ... μL DNA (see below)
A2: 12,6 μL
B1: 15,7 μL
B5: 9,4 μL
B6: 9,6 μL
D1: 10,2 μL
E1: 15,7 μL (1 μg)
I1: 14,3 μL (1 μg)
J1.2: 17,4 μL
CooA (8): 5,4 μL

Dephosphorylation + ligation using rAPId dephos and ligation protocol and kit

02

Preparing DNA for Submission

October 7

Preparing DNA for Submission

October 9

Dustin Vermeulen | Elise Grootscholten | Randall de Waard

Today we mini prepped our DNA (~6 colonies per succesfully sequenced biobrick, overnight cultures made on the 8th of october) and digested this.

03

Preparing DNA for Submission

October 9

Testing gas production

October 11

Elise Grootscholten | Paul Reusink

Overnight cultures made of the following strains with the right plasmid:

Number BBa_ HB101 JM109
J1 BBa_K2604000 1x 1x
I1 BBa_K2604001 3x 3x
D1 BBa_K2604003 2x 2x
B1 BBa_K2604006 1x 1x
B6 BBa_K2604006 1x 1x

Table 1: Overnight culture scheme.

I1, D1 and J1 are tested for the gas productio. The rest is only made for glycerolstocks

We can not say for sure if the PBad promoter and the gas production enzyme pyruvate decarboxylase work. Fot that more gas production test have to be done. Also the negative controls produce gas. This is a problem. The JM109 and HB101 E.coli strains didn't produce gas in experiment . 

Experiment 6: Testing gasproduction with our new biobricks.

Testing our new biobricks K260401 in JM109 and HB101. As a negative control we used the other new biobricks ...

Materials
-LB culture medium
-Kanamycin
-Bacteria on agar plates with the right plasmid

Methods
Make overnight cultures of the following strains with the right plasmid:
 

Number BBa_ HB101 JM109
J1 BBa_K2604000 1x 1x
I1 BBa_K2604001 3x 3x
D1 BBa_K2604003 2x 2x
B1 BBa_K2604006 1x 1x
B6 BBa_K2604006 1x 1x
Table 1: Overnight culture scheme.

Only the numbers J1, I1 and D1 are tested. The rest is made for making glycerolstocks.
J1 and D1 are the negative controls in this experiment.

LB: 582 mL +18 mL 100 mM sodiumpyruvate and 525 μL kanamycin (40 mg/mL).
After this the LB is devided in 2. Number 1: 0,5 μL arabinose, number 2: 1,0 μL arabinose.
The rest is done just like earlier experiments with a total of 20 tubes because everthing is done twice.

Results
Some of the I1 (BBa_K2604001) JM109 and HB101 bacteria and the negative controls D1 and J1 (BBa_K2604003 and BBa_K2604000) have produced gas over the weekend.
We can not say for sure if the PBad promoter and the gas production enzyme pyruvate decarboxylase work. Fot that more gas production test have to be done. Also the negative controls produce gas. This is a problem. The JM109 and HB101 E.coli strains didn't produce gas in experiment . 

Figure 1: Gas testing before the weekend, II1.1 HB101 nr.1/ I1.2 HB101 nr.1/ I1.3, HB101 nr.1/ I1.1 JM109 nr.1

Figure 2: Gas testing after the weekend, II1.1 HB101 nr.1/ I1.2 HB101 nr.1/ I1.3, HB101 nr.1/ I1.1 JM109 nr.1

Figure 3: Gas testing before the weekend, I1.3 JM109 nr.1/ I1.2 JM109 nr.1/ J1 JM109 nr.1/ D1 HB101 nr.1

Figure 4: Gas testing after the weekend, I1.3 JM109 nr.1/ I1.2 JM109 nr.1/ J1 JM109 nr.1/ D1 HB101 nr.1

Figure 5: Gas testing before the weekend, D1 JM109 nr.1/ J1 HB101 nr.1

Figure 6: Gas testing after the weekend, D1 JM109 nr.1/ J1 HB101 nr.1

Figure 7: Gas production before the weekend, I1.3 JM109 nr.2/ I1.2 JM109 nr.2/ I1.1 JM109 nr.2/ I1.3 HB101 nr.2

Figure 8: Gas production after the weekend, I1.3 JM109 nr.2/ I1.2 JM109 nr.2/ I1.1 JM109 nr.2/ I1.3 HB101 nr.2

Figure 9: Gas production before the weekend, D1 JM109 nr.2/ D1 HB101 nr.2/ J1 HB101 nr.2/ J1 JM109 nr.2

Figure 10: Gas production after the weekend, D1 JM109 nr.2/ D1 HB101 nr.2/ J1 HB101 nr.2/ J1 JM109 nr.2

Figure 11: Gas production before the weekend, I1.2 HB101 nr.2/ I1.1 HB101 nr.2

Figure 12: Gas production after the weekend, I1.2 HB101 nr.2/ I1.1 HB101 nr.2

06

Testing gas production

October 11

Assessing different ATP sensors

October 15

Loraine Nelson | Elise Grootscholten | Paul Reusink

Biobricks from iGEM were transformed into NEB10Beta. A few of these are sensors that react to ATP and can be used to detect living cells. The Bacteria were plated on agar plates with the right antibiotics to select the bacteria with the right plasmid in them. Every transformation was successful, since there were over 300 colonies on every plate. These colonies could than be used for further experiments and can be easily used for DNA isolation. 

Materials: 

- Biobricks 
- Nuclease free water
- LB- agar
- Variable Volume Pipettes 
- Sterile pipetpoints
- Chemically competent cells ( NEB10Beta, 1x10^9) 
- Heater (42°C)  
- Incubator + shaker (37°C and 250 rpm) 
- Eppendorf tubes (1,5 and 2 ml) 
- Stable outgrow medium for NEB10Beta or SOC- outgrow medium 

Method: 

To be able to use the DNA from the biobricks, they were first diluted in Nuclease free water. This was done by adding 10ul of nuclease free water to the DNA and incubated for 5 minutes at room temperature (18-24°C). After incubation the DNA is transferred to Eppendorf tubes and put on ice. 

For the transformation 1 ul of the DNA was added to 50 ul chemically competent cells strain NEB10Beta. The mixture was incubated for 30 minutes on ice. The mixture got a heat shock by putting the tube in a heater (42°C) for 30 seconds. After the heat shock the tube was immediately put on ice for 5 minutes. 
For good grow of the cells, 950 ul of outgrow medium was added to the mixture. After mixing the cells with outgrow medium, the culture was incubated for 1 hour at 37°C and 250 rpm.  

After incubation the culture was plated onto LB-agar plates. For specific growth of bacteria there was antibiotics added to the plates. The antibiotics that were used, depended on the plasmid. The biobricks used in this experiment were resistent to chloramphenicol (Can). We worked with a work concentration of 35 ug/ml for chloramphenicol. For good results we plated 100 ul undiluted culture onto a plate and centrifuged the rest of the culture. The supernatant was discarded and the pellet was resuspended and plated onto a LB-agar plate. The plates were then incubated by 37°C. 

Results:
Tabel 1 shows the results of the plates which were incubated. 

Biobricks:  Antibiotics:  Dilution:  Colonies: CFU: 
Bba-K352002 Can  0x  >300 -
Bba-K352002 Can Centrifuged >300 -
Bba-K352003 Can 0x >300 -
Bba-K352003 Can Centrifuged >300  -      
Bba-K352004 Can 0x >300 -
Bba-K352004 Can Centrifuged >300 -
Bba-K352005 Can 0x >300 -
Bba-K352005 Can Centrifuged >300 -
Bba-K352006 Can 0x >300 -
Bba-K352006 Can Centrifuged >300 -
Bba-K352007 Can 0x >300 -
Bba-K352007 Can Centrifuged >300 -
Bba-K352008 Can 0x >300 -
Bba-K352008 Can Centrifuged >300 -
Bba-K352009 Can 0x >300 -
Bba-K352009 Can Centrifuged >300 -
Bba-K1390001 Can 0x >300 -
Bba-K1390001 Can Centrifuged >300 -
Bba-K1390003 Can 0x >300 -
Bba-K1390003 Can Centrifuged >300 -
Bba-K1023003 Can 0x >300 -
Bba-K1023003 Can Centrifuged >300 -
Bba-K284001 amp 0x >300 -
Bba-K284001 amp Centrifuged >300 -
Positive controle: Bba- K1023003 None 0x >300 -   
Negative controle: outgrow medium none 0x 0 -

Tabel 1: results from the transformation with the biobricks. 

Showed in the Tabel, the transformation gave  enough colonies on each plate, which can be used for further experiments. There is no contamination, as seen by the negative control. 

Conclusion: 

The transformation from the biobricks was successful. The bacteria with the biobricks plasmids will be stored by 4°C and can be used for further experiments. 
 

04

Assessing different ATP sensors

October 15

Assessing different ATP sensors

October 15

Loraine Nelson | Elise Grootscholten | Paul Reusink

The bacteria were mini prepped and than transformed into BL21 DE3. BL21 DE3 has a T7 promotor which is needed for expression of the biobricks. The transformation was successful and multiple colonies were formed on the agar plates with antibiotics. From this plates we grow a few colonies onto new plates for further experiments. 

Transformation Protocol NEB
Overview

Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.

Protocol

  1. Thaw competent cells on ice.
  2. Chill approximately 5 ng (2 μl) of the ligation mixture in a 1.5 ml microcentrifuge tube.
  3. Add 50 μl of competent cells to the DNA. Mix gently by pipetting up and down or flicking the tube 4 to 5 times to mix the cells and DNA. Do not vortex.
  4. Place the mixture on ice for 30 minutes. Do not mix.
  5. Heat shock at 42°C for 30 seconds*. Do not mix.
  6. Add 950 μl of room temperature media* to the tube.
  7. Place tube at 37°C for 60 minutes. Shake vigorously (250 rpm) or rotate.
  8. Warm selection plates to 37°C.
  9. Spread 50 to100 μl of the cells and ligation mixture onto the plates.
  10. Incubate overnight at 37°C.

    * Please note: For the duration and temperature of the heat shock step as well as for the media to be used during the recovery period, please follow the recommendations provided by the competent cells� manufacturer.

    1 ul isolated K1023003 biobrick + 50 ul NEB10bèta 
    Positive and negative control without antibiotics

     

05

Assessing different ATP sensors

October 15

Assessing Gas Production BioBricks in E.Coli

October 16

Elise Grootscholten | Randall de Waard

Glycerolstocks have been made of several bacteria with plasmids we created or from the iGEM kit.

22

Assessing Gas Production BioBricks in E.Coli

October 16

Assessing CooA Production

October 16

Dustin van der Meulen

After performing a digestion on the 16th we ran a gel electrophoresis, from which we could conlcude that only the combinations of J23100+B0031, J23100+B0032 and K352001+B0015 seemed to be correct, though because of the small insert size in the promoter and rbs combinations it is difficult to say. From these combinations overnight cultures where made, which we prepped today. 

J23100+B0031 125,9 ng/uL
J23100+B0032 334,1 ng/μL
K352001+B0015 240,3 ng/μL

After this we've discussed our progress with our PI's, 

08

Assessing CooA Production

October 16

Chemo competent cells

October 16

None

New competent cells were made. This time we used the strains JM109 and HB101, since these strains don't produce gas of themselfes. These strains can be used for measurement on our gas production system. The cells are competent enough to use. 

Experiment 2: Chemocompetent cells of E.coli strains JM109 and HB101
28-09-18

Methods
The methods are for JM109 and HB101 both apart.
2 ul pUC19 + 50 ul competent cells
With ampicilin: not diluted, 10^-1, 10^-2, 10^-3, 10^-4
Without ampicillin: not diluted, 10^-4, 10^-5, 10^-6
Negative control: SOC medium.

200 ul not diluted -->
30 ul + 270 ul SOC (10^-1) --> 
30 ul + 270 ul SOC (10^-2) --> 
30 ul + 270 ul SOC (10^-3) --> 
30 ul + 270 ul SOC (10^-4) --> 
30 ul + 270 ul SOC (10^-5) --> 
30 ul + 270 ul SOC (10^-6)

Results
JM109: 3,4*10^6 cfu/ug DNA
HB101: 4,4*10^6 cfu/ug DNA

02

Chemo competent cells

October 16

Preparing DNA for Submission

October 17

Dustin Vermeulen | Elise Grootscholten | Randall de Waard

Today we set our control digest from the 9th of october on gel (the gel was inconclusive because of poor quality), after which we put ~500 ng of DNA on our 96 wells submission plate (both the pSB1C3 and pSB1K3 (sequenced) versions of our biobricks). This was then dried in a 50°C stove for around 2 hours (until all wells were drie).

04

Preparing DNA for Submission

October 17

Relevant pages

Results

Hardware

Software

Experiment