Difference between revisions of "Team:ZJUT-China/InterLab"

 
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{{ZJUT-China}}
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clear:both;
 +
}
 +
/*add extra space to page break with clear class*/
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .clear.extra_space {
 +
height: 30px;
 +
}
 +
 
 +
/* horizontal line to divide the page*/
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .line_divider {
 +
    border-top: 1px solid #c4baba;
 +
  margin: auto;
 +
  width: 98%;
 +
}
 +
 
 +
 
 +
 
 +
 
 +
/*support classes*/
 +
/**************************************************************************************************************************************************************************************************/
 +
 
 +
 
 +
/*Button  */
 +
/************************************************/
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .button_link {
 +
font-size: 130%;
 +
margin: 30px auto;
 +
text-align: center;
 +
}
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .button_link a  {
 +
background-color: #5bc7d8;
 +
color: #635d5d !important;
 +
font-weight: bold;
 +
margin: auto;
 +
text-decoration: none !important;
 +
padding: 10px 15px;
 +
}
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .button_link a:hover {
 +
background-color: #f8b732 !important;
 +
}
 +
 
 +
 
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight {
 +
padding: 15px 20px;
 +
}
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight p,
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight h1,
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight h2,
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight h3,
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight h4,
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight h5,
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight h6 {
 +
padding: 5px 15px;
 +
}
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_background {
 +
background-color: #e4dede;
 +
}
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_A_top {
 +
    border-top: 4px solid #5bc7d8;
 +
}
 +
 
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_A_full {
 +
    border: 4px solid #5bc7d8;
 +
}
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_B_top {
 +
    border-top: 4px solid #f8b732
 +
}
 +
 
 +
 
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .highlight.decoration_B_full {
 +
    border: 4px solid #f8b732;
 +
}
 +
 
 +
 
 +
 
 +
 
 +
/*mobile*/
 +
/**************************************************************************************************************************************************************************************************/
 +
 
 +
 
 +
/* 1800px  */
 +
/************************************************/
 +
@media only screen and (max-width: 1800px) {
 +
.igem_2018_team_content { width: 100%;}
 +
.igem_2018_team_menu {display:block;}
 +
}
 +
 
 +
/* 1400px  */
 +
/************************************************/
 +
@media only screen and (max-width: 1400px) {
 +
.igem_2018_team_menu .menu_item { font-size:100%;}
 +
.igem_2018_team_menu .submenu .submenu_item { font-size:90%;}
 +
.igem_2018_team_menu {display:block;}
 +
}
 +
 
 +
@media only screen and (max-width: 1001px) {
 +
.igem_2018_team_menu {display:block;}
 +
}
 +
 
 +
/* 1000px  */
 +
/************************************************/
 +
@media only screen and (max-width: 1000px) {
 +
.igem_2018_team_content {width:100%; margin-left:0px;}
 +
.igem_2018_team_menu {display:none; margin-top: 45px; min-width:50%; width:50%;}
 +
.igem_2018_team_mobile_bar {display:block;}
 +
.igem_2018_team_content .igem_2018_team_column_wrapper .column.full_size, .igem_2018_team_content .igem_2018_team_column_wrapper .column.two_thirds_size,.igem_2018_team_content .igem_2018_team_column_wrapper .column.third_size {width:96%; }
 +
 
 +
}
 +
 
 +
 
 +
@media only screen and (max-width: 500px) {
 +
.igem_2018_team_menu {min-width:100%; width:100%; }
 +
}
 +
 
 +
 
 +
/**************************************************************************************************************************************************************************************************/
 +
 
 +
/*my styles  */
 +
/************************************************/
 +
.bigbox { border: 4px solid #e4dede; padding: 2% !important; width: 92%!important;}
 +
img{
 +
  max-width: 100%;
 +
  max-height: 100%;
 +
  padding-top: 10px;
 +
  padding-left: 30px;
 +
}
 +
.homepage_img{
 +
  margin-top: 41px;
 
   text-align: center;
 
   text-align: center;
 +
  background: url(https://static.igem.org/mediawiki/2018/6/63/T--ZJUT-China--lab.jpg);
 +
  background-size:cover;
 +
  width: 100% ;
 +
  min-height: 800px;
 +
  height: auto;
 +
  clear: both;
 +
  background-repeat: no-repeat;
 +
  background-size: 100% 100%;
 +
  overflow: hidden;
 
}
 
}
.bluetable{
+
 
  background-color: #63B8FF
+
.container{
 +
padding:0 30px 0 30px;
 +
display: block;
 
}
 
}
.yellowable{
+
.homepage_img{
   background: #B8860B
+
   text-align: center;
 +
  padding:0 0 20px 0;
 
}
 
}
 +
/* 顶栏固定 */
 +
.topbar{
 +
  position: fixed;
 +
  top: 0px;
 +
  height: 51px;
 +
  width: 100%;
 +
  padding-top: 18px;
 +
}
 +
/* 下拉按钮样式 */
 +
.dropbkgd{
 +
    background-image: url(https://static.igem.org/mediawiki/2018/c/ca/T--ZJUT-China--topbar1.jpg);
 +
    min-height: 51px;
 +
    height: auto;
 +
    width: 100% ;
 +
    clear: both;
 +
    background-repeat: no-repeat;
 +
    background-size: 100% 100%;
 +
    overflow: hidden;
 +
  }
 +
.drop{
 +
  color: transparent;
 +
  font-size: 20px;
 +
  line-height: 51px;
 +
  width: 100%;
 +
  height: 100%;
 +
}
 +
 +
.dropposition{
 +
    position: absolute;
 +
    left: 15.2%;
 +
    width: 100%;
 +
    height: 51px;
 +
}
 +
.drop:hover{
 +
  background-color: #BACD32;
 +
  color: white;
 +
}
 +
.drop a{
 +
  color: transparent;
 +
  text-decoration: none;
 +
  display: inline-block;
 +
    width: 100%;
 +
    height: 100%;
 +
}
 +
.drop a:hover{
 +
    color: white;
 +
}
 +
 +
/* 容器 <div> - 需要定位下拉内容 */
 +
.dropdown1,
 +
.dropdown2,
 +
.dropdown3,
 +
.dropdown4,
 +
.dropdown5,
 +
.dropdown6,
 +
.dropdown7,
 +
.dropdown8{
 +
  position: relative;
 +
  display: inline-block;
 +
  float:left;
 +
  text-align: center;
 +
  width: 8%;
 +
  height: 100%;
 +
}
 +
.dropdown1 {
 +
    width: 7%;
 +
}
 +
.dropdown2{
 +
    width: 7%;
 +
    left: 2%;
 +
}
 +
.dropdown3{
 +
  width: 8%;
 +
  left: 3%;
 +
}
 +
.dropdown4{
 +
  width:7%;
 +
  left:5%;
 +
}
 +
.dropdown5{
 +
  width: 7%;
 +
  left:6%;
 +
}
 +
.dropdown6{
 +
  width:15.2%;
 +
  left: 7%;
 +
}
 +
.dropdown7{
 +
  width:12%;
 +
  left: 7%;
 +
}
 +
.dropdown8{
 +
  width:12%;
 +
  left: 8%;
 +
}
 +
/* 下拉内容 (默认隐藏) */
 +
.dropdown-content {
 +
    display: none;
 +
    position: absolute;
 +
    color: #DACD32;
 +
    width: 180px;
 +
    box-shadow: 0px 8px 16px 0px rgba(0,0,0,0.2);
 +
    float: left;
 +
    background-color: rgba(0,0,0,0.2);
 +
}
 +
.dropdown6 .dropdown-content{
 +
  width:230.4px;
 +
}
 +
 +
/* 下拉菜单的链接 */
 +
.dropdown-content a {
 +
    color: white;
 +
    text-decoration: none;
 +
    display: block;
 +
    width: 100%;
 +
    height: 100%;
 +
}
 +
 +
/* 鼠标移上去后修改下拉菜单链接颜色 */
 +
.dropdown-content a:hover {background-color:  #7D7DFF}
 +
 +
/* 在鼠标移上去后显示下拉菜单 */
 +
.dropdown2:hover .dropdown-content {
 +
    display: block;
 +
}
 +
.dropdown3:hover .dropdown-content {
 +
    display: block;
 +
}
 +
.dropdown4:hover .dropdown-content {
 +
    display: block;
 +
}
 +
.dropdown5:hover .dropdown-content {
 +
    display: block;
 +
}
 +
.dropdown6:hover .dropdown-content {
 +
    display: block;
 +
}
 +
.dropdown7:hover .dropdown-content {
 +
    display: block;
 +
}
 +
.dropdown8:hover .dropdown-content {
 +
    display: block;
 +
}
 +
 +
  .container .col-10 {
 +
    margin:auto;
 +
    max-width: 1400px;
 +
    width:88%;
 +
    padding: 20px 0 0 0 ;
 +
  }
 +
 +
  .container .col-10 h1 { font-size: 210%;}
 +
.container .col-10 h2 { font-size: 190%;}
 +
.container .col-10 h3 { font-size: 170%;}
 +
.container .col-10 h4 { font-size: 150%;}
 +
.container .col-10 h5 { font-size: 140%;}
 +
.container .col-10 h6 { font-size: 130%;}
 +
 +
 +
/* styling for the titles h1, h2*/
 +
  .container .col-10 h1, .container .col-10 h2 {
 +
border-bottom:0px;
 +
font-family: 'Tempus Sans ITC',georgia !important;
 +
padding: 10px 0px;
 +
}
 +
  .container .col-10 h1{
 +
    font-size: 38px!important;
 +
    padding: 20 20 20 20!important;
 +
    text-align: center;
 +
    color: #9ACD32;
 +
  }
 +
  .container .col-10 h2 {
 +
    font-size: 30px!important;
 +
    color: #6495ED;
 +
  }
 +
 +
 +
 +
/* styling for the titles h3, h3, h5, h6 */
 +
.container .col-10 h3,
 +
  .container .col-10 h4,
 +
.container .col-10 h5,
 +
.container .col-10 h6 {
 +
border-bottom:0px;
 +
color: #9ACD32;
 +
font-family: "Arial Black", Gadget, sans-serif;
 +
padding: 5px 0px;
 +
}
 +
 +
/* text */
 +
  .container .col-10 p {
 +
font-family: Arial, Helvetica, sans-serif;
 +
padding: 5px 0px;
 +
text-align: left;
 +
color: #000000;
 +
    font-size: 20px!important;
 +
}
 +
  .img-10{
 +
  text-align: center ;
 +
  weight: 600px;
 +
  }
 +
.side-bar .igem_2018_title_menu{
 +
    position: fixed;
 +
}
 +
.note{
 +
  font-size:15px;
 +
  color:#708090;
 +
  text-align: center;
 +
}
 +
.homeindex{
 +
  width: 20%;
 +
  min-height: 1px;
 +
  float: left;
 +
  margin: 15px;
 +
  text-align: center;
 +
  padding-left: 25px;
 +
 +
}
 +
.homeindex img{
 +
    border-radius: 30px;
 +
}
 +
/* 底部 */
 +
  .footer{
 +
    background: #eee;
 +
    margin-top: 30px;
 +
    padding: 30px 0;
 +
    width: 100%;
 +
  }
 +
  .foottop{
 +
    height: 330px;
 +
    width: 100%;
 +
  }
 +
  .footbottom{
 +
    height:50px;
 +
    width: 100%;
 +
  }
 +
  .footbottom .note{
 +
    padding-top: 40px;
 +
  }
 +
  .footmessage-left,
 +
  .footmessage-left2,
 +
  .footmessage-right,
 +
  .footmessage-middle {
 +
    width: 24%;
 +
    float: left;
 +
    height: 100%;
 +
  }
 +
  .footmessage-middle img{
 +
    width: 60%;
 +
    padding-top: 60px;
 +
  }
 +
  .footmessage-right img{
 +
    width: 80%;
 +
    max-height: 100%;
 +
    padding-top: 10px;
 +
    padding-left: 30px;
 +
  }
 +
  .footmessage-left2 img{
 +
    width: 80%;
 +
    padding-left: 30px;
 +
    padding-top: 15px;
 +
  }
 +
  .footmessage-left img{
 +
    padding-top: 60px;
 +
    padding-left: 30px;
 +
    width: 80%;
 +
    max-height: 100%;
 +
  }
 +
  .footmessage-right h3{
 +
    font-size: 30px;
 +
  }
 +
  .footmessage-right{
 +
    padding-left: 30px;
 +
  }
 +
  .footmessage-right p{
 +
    font-size: 15px!important;
 +
  }
 +
.container .igem_2018_team_content .igem_2018_team_column_wrapper  li{
 +
  padding-top: 10px;
 +
}
 +
.container .igem_2018_team_content .igem_2018_team_column_wrapper  a{
 +
  font-size: 20px;
 +
}
 +
.container .col-10 .igem_2018_team_content .igem_2018_team_column_wrapper h3{
 +
  text-align: center;
 +
}
 +
.col-2{
 +
  width: 45%;
 +
  float: left;
 +
  padding-left: 10px;
 +
  display: inline-block;
 +
}
 +
.clear{
 +
  clear: both;
 +
}
 +
.footmessage-middle img{
 +
      padding-left: 60px;
 +
        }
 
</style>
 
</style>
<!--interlab-->
+
 
<div class="column full_size judges-will-not-evaluate middle" >
+
 
<h3>★ OD600 reference point ★ </h3>
+
 
<br> <br>
+
 
<table border="1">
+
<!------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------->
  <thead>
+
<!--- THIS IS WHERE THE HTML BEGINS --->
  <tr>
+
<!------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------->
    <th></th>
+
 
    <th>LUDOX CL-X</th>
+
<head>
    <th>H2O</th>
+
 
  </tr>
+
<!-- This tells the browser that your page is responsive -->
</thead>
+
<meta name="viewport" content="width=device-width, initial-scale=1">
  <tbody>
+
 
  <tr>
+
</head>
    <th>Replicate 1</th>
+
<!-- 导航栏 -->
    <th class="bluetable">0.066</th>
+
<div class="topbar">
    <th class="bluetable">0.0285</th>
+
      <div class="dropbkgd">
  </tr>
+
        <div class="dropposition">
  <tr>
+
          <div class="dropdown1">
    <th>Replicate 2</th>
+
            <div class="drop">
    <th>0.059</th>
+
              <a href="https://2018.igem.org/Team:ZJUT-China">Home</a>
    <th>0.029</th>
+
            </div>
  </tr>
+
          </div>
  <tr>
+
          <div class="dropdown2">
    <th>Replicate 3</th>
+
            <div class="drop">
    <th>0.068</th>
+
                Team
    <th>0.026</th>
+
              <div class="dropdown-content">
  </tr>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Team">Team Members</a>
  <tr>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Attributions">Attributions</a>
    <th>Replicate 4</th>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Collaborations">Collaborations</a>
    <th>0.064</th>
+
              </div>
    <th>0.028</th>
+
            </div>
  </tr>
+
          </div>
  <tr>
+
          <div class="dropdown3">
    <th>Arith. Mean</th>
+
            <div class="drop">
    <th>0.064</th>
+
                Project
    <th>0.028</th>
+
              <div class="dropdown-content">
  </tr>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Description" >Description</a>
  <tr>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Design">Design</a>
    <th>Corrected Abs600</th>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Results">Results</a>
    <th> 0.036</th>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Demonstrate">Demonstrate</a>
    <th></th>
+
              </div>
  </tr>
+
            </div>
  <tr>
+
          </div>
    <th>Reference OD600</th>
+
          <div class="dropdown4">
    <th>OD600</th>
+
            <div class="drop">
    <th></th>
+
                Parts
  </tr>
+
              <div class="dropdown-content">
  <tr>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Parts">All parts</a>
    <th>OD600/Abs600</th>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Improve">Improve</a>
    <th>1.728</th>
+
              </div>
    <th></th>
+
            </div>
  </tr>
+
          </div>
  </tbody>
+
          <div class="dropdown5">
</table>
+
            <div class="drop">
</div>
+
              Lab
<div class="column full_size judges-will-not-evaluate middle" >
+
              <div class="dropdown-content">
  <h3>★ Fluorescence Raw Readings:★ </h3>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Notebook">Notebook</a>
  <br> <br>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Protocols2018">Protocols</a>
  <table border="1">
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Safety">Safety</a>
    <thead>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/InterLab" >Interlab</a>
    <tr>
+
              </div>
    <th>Hour 0:</th>
+
            </div>
    <th>Neg. Control</th>
+
          </div>
    <th>Pos. Control</th>
+
          <div class="dropdown6">
    <th>Device 1</th>
+
            <div class="drop">
    <th>Device 2</th>
+
                Human Practices
    <th>Device 3</th>
+
              <div class="dropdown-content">
    <th>Device 4</th>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Human_Practices">Integrated hp</a>
    <th>Device 5</th>
+
                <a href="https://2018.igem.org/Team:ZJUT-China/Public_Engagement">Public Engagement & Education</a>
    <th>Device 6</th>
+
              </div>
    <th>LB + Chlor (blank)</th>
+
            </div>
    </tr>
+
          </div>
    </thead>
+
          <div class="dropdown7">
    <tbody>
+
            <div class="drop">
    <tr>
+
              <a href="https://2018.igem.org/Team:ZJUT-China/Model"> Modeling</a>
    <th>Colony 1, Replicate 1</th>
+
            </div>
    <th>98.891</th>
+
          </div>
    <th>514.509</th>
+
          <div class="dropdown8">
    <th>170.45</th>
+
            <div class="drop">
    <th>453.211</th>
+
              <a href="https://2018.igem.org/Team:ZJUT-China/Achievement2018"> Judging</a>
    <th>107.527</th>
+
            </div>
    <th>377.998</th>
+
          </div>
    <th>110.335</th>
+
        </div>
    <th>121.276</th>
+
      </div>
    <th>100.56</th>
+
     </div>
    </tr>
+
<!-- 首页大图 -->
    <tr>
+
<div class="homepage_img">
    <th>Colony 1, Replicate 2</th>
+
    <th>87.951</th>
+
    <th>520.569</th>
+
    <th>155.638</th>
+
    <th>443.722</th>
+
    <th>103.159</th>
+
    <th>353.38</th>
+
    <th>99.569</th>
+
    <th>117.851</th>
+
    <th>97.983</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 3</th>
+
    <th>89.665</th>
+
    <th>500.61</th>
+
    <th>151.21</th>
+
    <th>474.651</th>
+
    <th>99.334</th>
+
    <th>358.843</th>
+
    <th>97.329</th>
+
    <th>115.674</th>
+
    <th>97.686</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 4</th>
+
    <th>88.651</th>
+
    <th>553.489</th>
+
    <th>158.106</th>
+
    <th>471.047</th>
+
    <th>107.222</th>
+
    <th>391.116</th>
+
    <th>102.558</th>
+
    <th>132.724</th>
+
    <th>104.901</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 1</th>
+
    <th>94.201</th>
+
    <th>799.161</th>
+
    <th>176.469</th>
+
    <th>396.183</th>
+
    <th>105.65</th>
+
    <th>452.456</th>
+
    <th>105.862</th>
+
    <th>129.379</th>
+
    <th>100.254</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 2</th>
+
    <th>97.455</th>
+
    <th>750.414</th>
+
    <th>168.71</th>
+
    <th>425.555</th>
+
    <th>106.286</th>
+
    <th>462.598</th>
+
    <th>109.024</th>
+
    <th>125.311</th>
+
    <th>101.914</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 3</th>
+
    <th>95.836</th>
+
    <th>795.354</th>
+
    <th>174.674</th>
+
    <th>405.952</th>
+
    <th>103.704</th>
+
    <th>444.673</th>
+
    <th>106.365</th>
+
    <th>128.606</th>
+
    <th>105.761</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 4</th>
+
    <th>100.733</th>
+
    <th>804.913</th>
+
    <th>203.631</th>
+
    <th>370.775</th>
+
    <th>117.227</th>
+
    <th>506.476</th>
+
    <th>113.511</th>
+
    <th>125.021</th>
+
    <th>105.18</th>
+
    </tr>
+
  </tbody>
+
  </table>
+
  <br> <br>
+
  <table border="1">
+
    <thead>
+
    <tr>
+
    <th>Hour 6:</th>
+
    <th>Neg. Control</th>
+
    <th>Pos. Control</th>
+
    <th>Device 1</th>
+
    <th>Device 2</th>
+
    <th>Device 3</th>
+
    <th>Device 4</th>
+
    <th>Device 5</th>
+
    <th>Device 6</th>
+
    <th>LB + Chlor (blank)</th>
+
    </tr>
+
    </thead>
+
    <tbody>
+
    <tr>
+
    <th>Colony 1, Replicate 1</th>
+
    <th>142.426</th>
+
    <th>1579.487</th>
+
    <th>407.651</th>
+
    <th>2268.46</th>
+
    <th>167.703</th>
+
    <th>1014.837</th>
+
    <th>144.06</th>
+
    <th>365.966</th>
+
    <th>130.779</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 2</th>
+
    <th>143.238</th>
+
    <th>1593.226</th>
+
    <th>396.943</th>
+
    <th>2328.22</th>
+
    <th>174.862</th>
+
    <th>1003.434</th>
+
    <th>141.231</th>
+
    <th>339.499</th>
+
    <th>126.638</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 3</th>
+
    <th>151.515</th>
+
    <th>1699.317</th>
+
    <th>396.669</th>
+
    <th>2175.501</th>
+
    <th>169.71</th>
+
    <th>977.6</th>
+
    <th>143.762</th>
+
    <th>347.711</th>
+
    <th>127.911</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 4</th>
+
    <th>155.799</th>
+
    <th>1783.066</th>
+
    <th>406.205</th>
+
    <th>2240.128</th>
+
    <th>171.295</th>
+
    <th>1044.087</th>
+
    <th>154.138</th>
+
    <th>340.887</th>
+
    <th>127.237</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 1</th>
+
    <th>150.46</th>
+
    <th>2062.959</th>
+
    <th>354.432</th>
+
    <th>2090.23</th>
+
    <th>165.189</th>
+
    <th>1268.176</th>
+
    <th>150.674</th>
+
    <th>320.936</th>
+
    <th>120.878</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 2</th>
+
    <th>145.579</th>
+
    <th>2023.17</th>
+
    <th>366.367</th>
+
    <th>2169.758</th>
+
    <th>167.611</th>
+
    <th>1203.787</th>
+
    <th>144.458</th>
+
    <th>322.957</th>
+
    <th>127.103</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 3</th>
+
    <th>146.553</th>
+
    <th>2101.747</th>
+
    <th>353.019</th>
+
    <th>1926.808</th>
+
    <th>183.724</th>
+
    <th>1172.553</th>
+
    <th>158.135</th>
+
    <th>332.951</th>
+
    <th>131.201</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 4</th>
+
    <th>157.503</th>
+
    <th>2071.459</th>
+
    <th>316.402</th>
+
    <th>1750.215</th>
+
    <th>179.275</th>
+
    <th>1253.449</th>
+
    <th>155.625</th>
+
    <th>327.385</th>
+
    <th>137.828</th>
+
    </tr>
+
  </tbody>
+
  </table>
+
</div>
+
<div class="column full_size judges-will-not-evaluate middle" >
+
  <h3>★ Abs600 Raw Readings:★ </h3>
+
  <br> <br>
+
  <table border="1">
+
    <thead>
+
    <tr>
+
    <th>Hour 0:</th>
+
    <th>Neg. Control</th>
+
    <th>Pos. Control</th>
+
    <th>Device 1</th>
+
    <th>Device 2</th>
+
    <th>Device 3</th>
+
    <th>Device 4</th>
+
    <th>Device 5</th>
+
    <th>Device 6</th>
+
    <th>LB + Chlor (blank)</th>
+
    </tr>
+
    </thead>
+
    <tbody>
+
    <tr>
+
    <th>Colony 1, Replicate 1</th>
+
    <th>0.1306</th>
+
    <th>0.0884</th>
+
    <th>0.0767</th>
+
    <th>0.1068</th>
+
    <th>0.1181</th>
+
    <th>0.0753</th>
+
    <th>0.1067</th>
+
    <th>0.0866</th>
+
    <th>0.0486</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 2</th>
+
    <th>0.1272</th>
+
    <th>0.0906</th>
+
    <th>0.0737</th>
+
    <th>0.1075</th>
+
    <th>0.1184</th>
+
    <th>0.0754</th>
+
    <th>0.108</th>
+
    <th>0.0896</th>
+
    <th>0.0497</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 3</th>
+
    <th>0.1246</th>
+
    <th>0.09</th>
+
    <th>0.0747</th>
+
    <th>0.1119</th>
+
    <th>0.1214</th>
+
    <th>0.0731</th>
+
    <th>0.1083</th>
+
    <th>0.0879</th>
+
    <th>0.0464</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 4</th>
+
    <th>0.1225</th>
+
    <th>0.0872</th>
+
    <th>0.0743</th>
+
    <th>0.1109</th>
+
    <th>0.1247</th>
+
    <th>0.0746</th>
+
    <th>0.1079</th>
+
    <th>0.0903</th>
+
    <th>0.0476</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 1</th>
+
    <th>0.1301</th>
+
    <th>0.0907</th>
+
    <th>0.0701</th>
+
    <th>0.1109</th>
+
    <th>0.1132</th>
+
    <th>0.0813</th>
+
    <th>0.1103</th>
+
    <th>0.0909</th>
+
    <th>0.0468</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 2</th>
+
    <th>0.1327</th>
+
    <th>0.0919</th>
+
    <th>0.0685</th>
+
    <th>0.1125</th>
+
    <th>0.1145</th>
+
    <th>0.0822</th>
+
    <th>0.1081</th>
+
    <th>0.0896</th>
+
    <th>0.0472</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 3</th>
+
    <th>0.1294</th>
+
    <th>0.0952</th>
+
    <th>0.0693</th>
+
    <th>0.1151</th>
+
    <th>0.1139</th>
+
    <th>0.0809</th>
+
    <th>0.1084</th>
+
    <th>0.0898</th>
+
    <th>0.05</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 4</th>
+
    <th>0.1202</th>
+
    <th>0.0941</th>
+
    <th>0.0692</th>
+
    <th>0.1105</th>
+
    <th>0.1134</th>
+
    <th>0.0805</th>
+
    <th>0.1092</th>
+
    <th>0.089</th>
+
    <th>0.0472</th>
+
    </tr>
+
  </tbody>
+
  </table>
+
  <br> <br>
+
  <table border="1">
+
    <thead>
+
    <tr>
+
    <th>Hour 6:</th>
+
    <th>Neg. Control</th>
+
    <th>Pos. Control</th>
+
    <th>Device 1</th>
+
    <th>Device 2</th>
+
    <th>Device 3</th>
+
    <th>Device 4</th>
+
    <th>Device 5</th>
+
    <th>Device 6</th>
+
    <th>LB + Chlor (blank)</th>
+
    </tr>
+
    </thead>
+
    <tbody>
+
    <tr>
+
    <th>Colony 1, Replicate 1</th>
+
    <th>0.9386</th>
+
    <th>0.7564</th>
+
    <th>0.1978</th>
+
    <th>0.911</th>
+
    <th>0.7963</th>
+
    <th>0.2566</th>
+
    <th>0.8131</th>
+
     <th>0.6363</th>
+
    <th>0.0444</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 2</th>
+
    <th>0.9413</th>
+
    <th>0.7528</th>
+
    <th>0.1974</th>
+
    <th>0.9012</th>
+
    <th>0.8211</th>
+
    <th>0.2547</th>
+
    <th>0.7951</th>
+
    <th>0.5996</th>
+
    <th>0.0438</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 3</th>
+
    <th>0.9745</th>
+
    <th>0.7748</th>
+
    <th>0.1951</th>
+
    <th>0.8642</th>
+
    <th>0.81</th>
+
    <th>0.2536</th>
+
    <th>0.7994</th>
+
    <th>0.6044</th>
+
    <th>0.0458</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 1, Replicate 4</th>
+
    <th>0.9932</th>
+
    <th>0.7975</th>
+
    <th>0.2009</th>
+
    <th>0.8969</th>
+
    <th>0.8021</th>
+
    <th>0.2622</th>
+
    <th>0.8266</th>
+
    <th>0.6092</th>
+
    <th>0.0438</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 1</th>
+
    <th>0.8246</th>
+
    <th>0.78</th>
+
    <th>0.1031</th>
+
    <th>0.8625</th>
+
    <th>0.8193</th>
+
    <th>0.2834</th>
+
    <th>0.7633</th>
+
    <th>0.5251</th>
+
    <th>0.0457</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 2</th>
+
    <th>0.8327</th>
+
    <th>0.7665</th>
+
    <th>0.1031</th>
+
    <th>0.9018</th>
+
    <th>0.8211</th>
+
    <th>0.2781</th>
+
    <th>0.7394</th>
+
    <th>0.5374</th>
+
    <th>0.0462</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 3</th>
+
    <th>0.8062</th>
+
    <th>0.7927</th>
+
    <th>0.1005</th>
+
    <th>0.829</th>
+
    <th>0.8082</th>
+
    <th>0.276</th>
+
    <th>0.7689</th>
+
    <th>0.5668</th>
+
    <th>0.0458</th>
+
    </tr>
+
    <tr>
+
    <th>Colony 2, Replicate 4</th>
+
    <th>0.8262</th>
+
    <th>0.7805</th>
+
    <th>0.1005</th>
+
    <th>0.8787</th>
+
    <th>0.8156</th>
+
    <th>0.2928</th>
+
    <th>0.7604</th>
+
    <th>0.5332</th>
+
    <th>0.0477</th>
+
    </tr>
+
  </tbody>
+
  </table>
+
 
</div>
 
</div>
<div class="column full_size judges-will-not-evaluate middle" >
+
<!-- 主体 -->
  <h3>★ Assumed plate well pattern:★ </h3>
+
<div class="container">
  <br> <br>
+
  <div class="col-10">
  <table border="1">
+
    <div class="igem_2018_team_content">
  <tr>
+
      <div class="igem_2018_team_column_wrapper">
    <th>A1</th>
+
        <div class="clear extra_space"></div>
    <th>A2</th>
+
        <div class="clear extra_space"></div>
    <th>A3</th>
+
      <!--interlab-->
    <th>A4</th>
+
        <div class="column full_size bigbox ">
    <th>A5</th>
+
          <h1>Interlab</h1>
    <th>A6</th>
+
          <p>Our team has participated in the Fifth International InterLaboratory Measurement Study this year. The Interlab study aims on solving the problem of reliability and repeatability of synthetic biology study. This year we were asked to measure green fluorescent protein, which is usually used as a measurement marker. We followed the experiment protocol strictly to make sure our data are valid. After two weeks' lab work, we got the data we needed after several attempts.</p>
    <th>A7</th>
+
        </div>
    <th>A8</th>
+
        <div class="column  bigbox middle" >
    <th>A9</th>
+
        <h1>Calibration Protocol</h1>
  </tr>
+
        <p>Three calibration measurements were done by following the calibration protocol, including an OD reference point at 600nm, one particle standard curve and one fluorescence standard curve.</p>
  <tr>
+
        <div class="note" style="color:red">(Plate Reader: Molecular Device, Spectramax M5)</div>
    <th>B1</th>
+
        <br><br>
    <th>B2</th>
+
        <h3>★ OD<sub>600</sub> reference point ★ </h3>
    <th>B3</th>
+
        <br>
    <th>B4</th>
+
        <table border="1">
    <th>B5</th>
+
          <thead>
    <th>B6</th>
+
            <tr>
    <th>B7</th>
+
              <th></th>
    <th>B8</th>
+
              <th>LUDOX CL-X</th>
    <th>B9</th>
+
              <th>H<sub>2</sub>O</th>
  </tr> <tr>
+
            </tr>
    <th>C1</th>
+
          </thead>
    <th>C2</th>
+
          <tbody>
    <th>C3</th>
+
            <tr>
    <th>C4</th>
+
              <th>Replicate 1</th>
    <th>C5</th>
+
              <th class="bluetable">0.066</th>
    <th>C6</th>
+
              <th class="bluetable">0.0285</th>
    <th>C7</th>
+
            </tr>
    <th>C8</th>
+
              <tr>
    <th>C9</th>
+
              <th>Replicate 2</th>
  </tr><tr>
+
              <th>0.059</th>
    <th>D1</th>
+
              <th>0.029</th>
    <th>D2</th>
+
            </tr>
    <th>D3</th>
+
            <tr>
    <th>D4</th>
+
              <th>Replicate 3</th>
    <th>D5</th>
+
              <th>0.068</th>
    <th>D6</th>
+
              <th>0.026</th>
    <th>D7</th>
+
            </tr>
    <th>D8</th>
+
            <tr>
    <th>D9</th>
+
              <th>Replicate 4</th>
  </tr><tr>
+
              <th>0.064</th>
    <th>E1</th>
+
              <th>0.028</th>
    <th>E2</th>
+
            </tr>
    <th>E3</th>
+
            <tr>
    <th>E4</th>
+
              <th>Arith. Mean</th>
    <th>E5</th>
+
              <th>0.064</th>
    <th>E6</th>
+
              <th>0.028</th>
    <th>E7</th>
+
            </tr>
    <th>E8</th>
+
            <tr>
    <th>E9</th>
+
              <th>Corrected Abs600</th>
  </tr><tr>
+
              <th> 0.036</th>
    <th>F1</th>
+
              <th></th>
    <th>F2</th>
+
            </tr>
    <th>F3</th>
+
            <tr>
    <th>F4</th>
+
              <th>Reference OD<sub>600</sub></th>
    <th>F5</th>
+
              <th>OD<sub>600</sub></th>
    <th>F6</th>
+
              <th></th>
    <th>F7</th>
+
            </tr>
    <th>F8</th>
+
            <tr>
    <th>F9</th>
+
              <th>OD<sub>600</sub>/Abs600</th>
  </tr><tr>
+
              <th>1.728</th>
    <th>G1</th>
+
              <th></th>
    <th>G2</th>
+
            </tr>
    <th>G3</th>
+
            </tbody>
    <th>G4</th>
+
        </table>
    <th>G5</th>
+
          <div class="note" style="color:red">Form1.OD reference point</div>
    <th>G6</th>
+
          <br><br>
    <th>G7</th>
+
            <div class="col-2">
    <th>G8</th>
+
              <img src="https://static.igem.org/mediawiki/2018/4/42/T--ZJUT-China--Particle_Standard_Curve.png"  alt="Particle_Standard_Curve.png">
    <th>G9</th>
+
              <div class="note" style="color:red">Fig1.Particle standard curve</div>
  </tr><tr>
+
            </div>
    <th>H1</th>
+
            <div class="col-2">
    <th>H2</th>
+
              <img src="https://static.igem.org/mediawiki/2018/f/fc/T--ZJUT-China--Particle_Standard_Curve_%28log_scale%29e.png" height="259.25px" alt="Particle_Standard_Curve_(log_scale)e">
    <th>H3</th>
+
              <div class="note" style="color:red">Fig2.Particle standard curve (log_scale)</div>
    <th>H4</th>
+
            </div>
    <th>H5</th>
+
          <div class="col-2">
    <th>H6</th>
+
            <img src="https://static.igem.org/mediawiki/2018/c/c5/T--ZJUT-China--Fluorescein_Standard_Curve.png" alt="Fluorescein_Standard_Curve">
    <th>H7</th>
+
            <div class="note" style="color:red">Fig3.Fluorescein standard curve</div>
    <th>H8</th>
+
          </div>
    <th>H9</th>
+
          <div class="col-2">
  </tr>
+
            <img src="https://static.igem.org/mediawiki/2018/c/ca/T--ZJUT-China--Fluorescein_Standard_Curve_%28log_scale%29.png" alt="Fluorescein_Standard_Curve_%28log_scale%29">
   </table>
+
            <div class="note" style="color:red">Fig4.Fluorescein standard curve (log_scale)</div>
 +
          </div>
 +
      </div>
 +
      <div class="column bigbox middle" >
 +
        <h1>Cell measurement</h1>
 +
        <h2>Transformation</h2>
 +
        <p>Transform <i>Escherichia coli</i> DH5α with these following plasmids:</p>
 +
        <table>
 +
          <tr>
 +
            <th>Device</th>
 +
            <th>Part Number</th>
 +
            <th>Location</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Positive control</th>
 +
            <th>BBa_R0040</th>
 +
            <th>Well 2D</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Negative control</th>
 +
            <th>Ba_I20270</th>
 +
            <th>Well 2B</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Test Device 1</th>
 +
            <th>BBa_J364000</th>
 +
            <th>Well 2F</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Test Device 2</th>
 +
            <th>BBa_J364001</th>
 +
            <th>Well 2H</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Test Device 3</th>
 +
            <th>BBa_J364002</th>
 +
            <th>Well 2J</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Test Device 4</th>
 +
            <th>BBa_J364007</th>
 +
            <th>Well 2L</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Test Device 5</th>
 +
            <th>BBa_J364008</th>
 +
            <th>Well 2N</th>
 +
          </tr>
 +
          <tr>
 +
            <th>Test Device 6</th>
 +
            <th>BBa_J364009</th>
 +
            <th>Well 2P</th>
 +
          </tr>
 +
        </table>
 +
        <div class="note" style="color:red">Form2.Devices (from Distribution Kit, all in pSB1C3 backbone) </div>
 +
        <p>The transformation used <i>E.coli</i> DH5α competent cells was bought from Tsingke Biological Technology company, and we followed the steps in http://parts.igem.org/Help:2017_DNA_Distribution to use the DNA in the Distribution Kit.</p>
 +
        <h2>Measurement </h2>
 +
        <br>
 +
        <h3>For this part of inter lab study, we followed the following protocols:
 +
          <br></h3>
 +
        <p>1.Grown 8 devices in incubator for 12 hrs at 37 ℃
 +
          <br>
 +
          2.Pick 2 colonies from each of plate and inocubate them on 10mL LB medium with Chloramphenicol. Grow the cells for 16-20hrs at 37°C and 180rpm.
 +
          <br>
 +
          3.Measure OD<sub>600</sub>nm of the overnight cultures and record the data, then dilute to target OD<sub></sub>nm = 0.02 in the falcon tubes.
 +
          <br>
 +
          4.Measure the OD<sub>600</sub>nm and Fl under the same condition as standard curve measurement and use the same 96 wells plates.</p>
 +
        <br>
 +
        <img src="https://static.igem.org/mediawiki/2018/5/5b/T--ZJUT-China--plate_lay_out.png" alt="plate_lay_out">
 +
        <br><div class="note" style="color:red">Fig5. plate lay out </div>
 +
        <br><br>
 +
        <div class="column bigbox middle" >
 +
        <h3>★ Fluorescence Raw Readings:★</h3>
 +
        <img src="https://static.igem.org/mediawiki/2018/d/d2/T--ZJUT-China--Fluorescence_Raw_Readings.png" alt="Fluorescence Raw Readings">
 +
        <br><br>
 +
        <h3>★ Abs600 Raw Readings:★ </h3>
 +
        <img src="https://static.igem.org/mediawiki/2018/c/ce/T--ZJUT-China--Abs600_Raw_Readings.png" alt="Abs600 Raw Readings">
 +
        <br><br>
 +
        <h3>★ uM Fluorescein / OD ★ </h3>
 +
        <img src="https://static.igem.org/mediawiki/2018/b/b6/T--ZJUT-China--11.png" alt="uM Fluorescein / OD">
 +
        <br><br>
 +
        <h3>★ Net Fluorescein a.u. ★ </h3>
 +
        <img src="https://static.igem.org/mediawiki/2018/5/5a/T--ZJUT-China--Net_Fluoresceina.u.png" alt="Net_Fluoresceina.u.">
 +
        <br><br>
 +
        <h3>★ Net Abs 600 ★ </h3>
 +
        <img src="https://static.igem.org/mediawiki/2018/5/55/T--ZJUT-China--Net_Abs_600.png" alt="Net_Abs_600">
 +
        <br><div class="note" style="color:red">Form3-6. Result of Raw Plate Reader Measurements </div>
 +
        </div>
 +
      </div>
 +
      <div class="column full_size bigbox middle">
 +
        <h1>CFU Protocol</h1>
 +
        <h3>For this part of inter lab study, we followed the following protocol:</h3>
 +
        <p>1.Pick two colonies from positive control device and negative control device, incubate overnight.
 +
          <br>
 +
          2.Prepare Starting Samples: Dilute the overnight cultures 1:8, measurement then dilute further to OD<sub>600</sub>nm = 0.1
 +
          <br>
 +
          Calculation:
 +
          <br>
 +
          Use (C1)(V1) = (C2)(V2) to calculate your dilutions
 +
          C1 is your starting OD600 &#8195; C2 is your target OD<sub>600</sub> of 0.1
 +
          <br>
 +
          V1 is the unknown volume in μL  &#8195;      V2 is the final volume of 1000 μL
 +
          <br>
 +
          3.Check and make sure OD<sub>600</sub> = 0.1
 +
          <br>
 +
          4.Dilute
 +
          <img src="https://static.igem.org/mediawiki/2018/e/ee/T--ZJUT-China--Dilute.png" alt="Dilute">
 +
          <br>
 +
          5.Incubate samples at 37 degree Celsius overnight for 18-20 hrs. Then count colony numbers.
 +
          <br>
 +
          <img src="https://static.igem.org/mediawiki/2018/4/4c/T--ZJUT-China--colony_number.png"  width="900px"alt="colony_number">
 +
        </p>
 +
      </div>
 +
      <div class="column full_size bigbox middle">
 +
        <h1>Disscussion & Feedback</h1>
 +
        <p>After the lab work, we had a discussion about the interlab study we did this year and now we have some feedbacks.
 +
        <br>    1.The interlab study protocol is very detailed and clear, we followed almost every step. But in some attempts, we tried to dilute the cultures in 96 wells plate, which is easier and quicker. So we do think improvements could be made to adjust the protocol.
 +
        <br>    2.The Excel sheets provide us an easy way to process our data. However, we still wish to know the meaning behind every function. It took us some time to search online, things would be easier if there is a guide.
 +
        <br>    3.The protocol tells us some principles of the experiment and it’s unbelievably helpful for us to complete the whole lnterlab study.
 +
        <br>    4.The kit needs a handbook to help those teams who participate in interlab study for the first time.
 +
        </p>
 +
      </div>
 +
      <div class="column full_size bigbox middle">
 +
        <h1>Collaboration</h1>
 +
        <p>
 +
        On August 8, 2018. We had a meeting with team jiangnan_China, team DLUT-China and team LZU-China in Jiangsu Normal University. By then we have already finished the interlab study successfully, but team jiangnan_China and team LZU-China met some difficulties. We provided them with some guidance and advice.
 +
        </p>
 +
      </div>
 +
      </div>
 +
    </div>
 +
   </div>
 
</div>
 
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      <div class="footmessage-right">
 +
        <h3>Contact</h3>
 +
        <p>
 +
        <br><br>
 +
        College of Biotechnology and Bioengineering
 +
        <br><br>Zhejiang University of Technology
 +
        <br><br>Chaowang Rd. 18, 310014, Hangzhou, China
 +
        <br><br>E-mail: cbb@zjut.edu.cn  Tel: +86-571-88320391
 +
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 +
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      Copyright © 2018 ZJUT-China
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Latest revision as of 16:24, 16 October 2018

Team:ZJUT-China - 2018.igem.org

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Team:ZJUT-China

Interlab

Our team has participated in the Fifth International InterLaboratory Measurement Study this year. The Interlab study aims on solving the problem of reliability and repeatability of synthetic biology study. This year we were asked to measure green fluorescent protein, which is usually used as a measurement marker. We followed the experiment protocol strictly to make sure our data are valid. After two weeks' lab work, we got the data we needed after several attempts.

Calibration Protocol

Three calibration measurements were done by following the calibration protocol, including an OD reference point at 600nm, one particle standard curve and one fluorescence standard curve.

(Plate Reader: Molecular Device, Spectramax M5)


★ OD600 reference point ★


LUDOX CL-X H2O
Replicate 1 0.066 0.0285
Replicate 2 0.059 0.029
Replicate 3 0.068 0.026
Replicate 4 0.064 0.028
Arith. Mean 0.064 0.028
Corrected Abs600 0.036
Reference OD600 OD600
OD600/Abs600 1.728
Form1.OD reference point


Particle_Standard_Curve.png
Fig1.Particle standard curve
Particle_Standard_Curve_(log_scale)e
Fig2.Particle standard curve (log_scale)
Fluorescein_Standard_Curve
Fig3.Fluorescein standard curve
Fluorescein_Standard_Curve_%28log_scale%29
Fig4.Fluorescein standard curve (log_scale)

Cell measurement

Transformation

Transform Escherichia coli DH5α with these following plasmids:

Device Part Number Location
Positive control BBa_R0040 Well 2D
Negative control Ba_I20270 Well 2B
Test Device 1 BBa_J364000 Well 2F
Test Device 2 BBa_J364001 Well 2H
Test Device 3 BBa_J364002 Well 2J
Test Device 4 BBa_J364007 Well 2L
Test Device 5 BBa_J364008 Well 2N
Test Device 6 BBa_J364009 Well 2P
Form2.Devices (from Distribution Kit, all in pSB1C3 backbone)

The transformation used E.coli DH5α competent cells was bought from Tsingke Biological Technology company, and we followed the steps in http://parts.igem.org/Help:2017_DNA_Distribution to use the DNA in the Distribution Kit.

Measurement


For this part of inter lab study, we followed the following protocols:

1.Grown 8 devices in incubator for 12 hrs at 37 ℃
2.Pick 2 colonies from each of plate and inocubate them on 10mL LB medium with Chloramphenicol. Grow the cells for 16-20hrs at 37°C and 180rpm.
3.Measure OD600nm of the overnight cultures and record the data, then dilute to target ODnm = 0.02 in the falcon tubes.
4.Measure the OD600nm and Fl under the same condition as standard curve measurement and use the same 96 wells plates.


plate_lay_out
Fig5. plate lay out


★ Fluorescence Raw Readings:★

Fluorescence Raw Readings

★ Abs600 Raw Readings:★

Abs600 Raw Readings

★ uM Fluorescein / OD ★

uM Fluorescein / OD

★ Net Fluorescein a.u. ★

Net_Fluoresceina.u.

★ Net Abs 600 ★

Net_Abs_600
Form3-6. Result of Raw Plate Reader Measurements

CFU Protocol

For this part of inter lab study, we followed the following protocol:

1.Pick two colonies from positive control device and negative control device, incubate overnight.
2.Prepare Starting Samples: Dilute the overnight cultures 1:8, measurement then dilute further to OD600nm = 0.1
Calculation:
Use (C1)(V1) = (C2)(V2) to calculate your dilutions C1 is your starting OD600   C2 is your target OD600 of 0.1
V1 is the unknown volume in μL   V2 is the final volume of 1000 μL
3.Check and make sure OD600 = 0.1
4.Dilute Dilute
5.Incubate samples at 37 degree Celsius overnight for 18-20 hrs. Then count colony numbers.
colony_number

Disscussion & Feedback

After the lab work, we had a discussion about the interlab study we did this year and now we have some feedbacks.
1.The interlab study protocol is very detailed and clear, we followed almost every step. But in some attempts, we tried to dilute the cultures in 96 wells plate, which is easier and quicker. So we do think improvements could be made to adjust the protocol.
2.The Excel sheets provide us an easy way to process our data. However, we still wish to know the meaning behind every function. It took us some time to search online, things would be easier if there is a guide.
3.The protocol tells us some principles of the experiment and it’s unbelievably helpful for us to complete the whole lnterlab study.
4.The kit needs a handbook to help those teams who participate in interlab study for the first time.

Collaboration

On August 8, 2018. We had a meeting with team jiangnan_China, team DLUT-China and team LZU-China in Jiangsu Normal University. By then we have already finished the interlab study successfully, but team jiangnan_China and team LZU-China met some difficulties. We provided them with some guidance and advice.