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body{ | body{ | ||
− | background: | + | background: white !important; |
− | font-family: | + | font-family: Levenim MT; |
} | } | ||
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margin-bottom: 2vh; | margin-bottom: 2vh; | ||
margin-top:3vh; | margin-top:3vh; | ||
− | |||
text-shadow: 0 1px 2px rgba(0,0,0,0.3); | text-shadow: 0 1px 2px rgba(0,0,0,0.3); | ||
− | margin-left: | + | width: 60%; |
+ | text-align: center; | ||
+ | margin-left: 20%; | ||
} | } | ||
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color: #053B77; | color: #053B77; | ||
background: #053B77; | background: #053B77; | ||
− | border: solid | + | border: solid 0.5px #053B77; |
+ | margin-top: 0.75em; | ||
+ | margin-bottom: 1em; | ||
} | } | ||
/*-------------------------- | /*-------------------------- | ||
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float:left; | float:left; | ||
margin-right:5px; | margin-right:5px; | ||
− | font-size: | + | font-size: 2.7vw; |
margin-bottom:-2px; | margin-bottom:-2px; | ||
font-weight:bold !important; | font-weight:bold !important; | ||
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.menu__tabs a { | .menu__tabs a { | ||
padding: 3vmin; | padding: 3vmin; | ||
− | padding-top: | + | padding-top: 3.5vmin; |
− | padding-bottom: | + | padding-bottom: 2vmin; |
font-weight:bold !important; | font-weight:bold !important; | ||
background: #b1e0f2; | background: #b1e0f2; | ||
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---------------------------*/ | ---------------------------*/ | ||
.menu { | .menu { | ||
− | margin-left: | + | margin-left:10%; |
− | + | margin-top:3vw; | |
width: 35%; | width: 35%; | ||
− | |||
position: relative; | position: relative; | ||
font-family: 'Myriad Pro'; | font-family: 'Myriad Pro'; | ||
+ | display: inline-block; | ||
+ | float: left; | ||
} | } | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | } | + | @media screen and( max-width:768px){ |
+ | .menu { | ||
+ | margin-left:10%; | ||
+ | margin-top:130px; | ||
+ | width: 80%; | ||
+ | position: relative; | ||
+ | font-family: 'Myriad Pro'; | ||
+ | } | ||
+ | .notebook { | ||
+ | width: 80%; | ||
+ | margin-left: 10%; | ||
+ | position: relative; | ||
+ | margin-top: 2vw; | ||
+ | } | ||
} | } | ||
.menu__wrapper { | .menu__wrapper { | ||
padding: 6vmin; | padding: 6vmin; | ||
padding-top: 4vmin; | padding-top: 4vmin; | ||
− | padding-left: | + | padding-left: 3vw; |
− | padding-bottom: | + | padding-bottom: 2vw; |
position: relative; | position: relative; | ||
z-index: 400; | z-index: 400; | ||
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th{ | th{ | ||
− | font-size:2. | + | font-size:2.3vw; |
text-align:center; | text-align:center; | ||
margin: 15px; | margin: 15px; | ||
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table tr td{ | table tr td{ | ||
− | font-size:2. | + | font-size:2.6vw; |
text-align:center; | text-align:center; | ||
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.notebook{ | .notebook{ | ||
− | margin-left: | + | margin-left:10%; |
− | margin- | + | margin-bottom:150px; |
− | margin- | + | width: 35%; |
+ | display: inline-block; | ||
+ | margin-top: 3vw; | ||
} | } | ||
.notebook_wrapper{ | .notebook_wrapper{ | ||
background:#b1e0f2; | background:#b1e0f2; | ||
− | width: | + | width:100%; |
height:490px; | height:490px; | ||
border-radius:10px; | border-radius:10px; | ||
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text-align:center; | text-align:center; | ||
color: #b1e0f2; | color: #b1e0f2; | ||
− | font-size: | + | font-size:4em; |
− | padding: | + | padding: 13px; |
border-radius: 10px; | border-radius: 10px; | ||
background: #053B77; | background: #053B77; | ||
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.notebook_wrapper .note-item .note-title{ | .notebook_wrapper .note-item .note-title{ | ||
− | font-size: | + | font-size:3em !important; |
font-weight:bold !important; | font-weight:bold !important; | ||
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padding-bottom:8px; | padding-bottom:8px; | ||
padding-left:5px !important; | padding-left:5px !important; | ||
+ | } | ||
+ | |||
+ | .lab{ | ||
+ | width: 15%; | ||
+ | position: absolute; | ||
+ | left: 73%; | ||
+ | top: 30vw; | ||
+ | transition-duration: 0.4s; | ||
+ | } | ||
+ | |||
+ | .protocol{ | ||
+ | width: 15%; | ||
+ | position: absolute; | ||
+ | left: 73%; | ||
+ | top: 37vw; | ||
+ | transition-duration: 0.4s; | ||
+ | } | ||
+ | |||
+ | .lab:hover{ | ||
+ | transform: scale(1.1); | ||
+ | } | ||
+ | |||
+ | .protocol:hover{ | ||
+ | transform: scale(1.1); | ||
} | } | ||
</style> | </style> | ||
</head> | </head> | ||
+ | |||
<body> | <body> | ||
<div class="img-container"> | <div class="img-container"> | ||
<img src="https://static.igem.org/mediawiki/2018/3/31/T--NCTU_Formosa--Notebook_cover.png" width="100%"> | <img src="https://static.igem.org/mediawiki/2018/3/31/T--NCTU_Formosa--Notebook_cover.png" width="100%"> | ||
+ | <a href="https://2018.igem.org/Team:NCTU_Formosa/Notebook"><img src="https://static.igem.org/mediawiki/2018/4/4c/T--NCTU_Formosa--Notebook_Lab_Note.png" class="lab"></a> | ||
+ | <a href="https://2018.igem.org/Team:NCTU_Formosa/Protocol"><img src="https://static.igem.org/mediawiki/2018/4/4f/T--NCTU_Formosa--Notebook_Lab_protocol.png" class="protocol"></a> | ||
</div> | </div> | ||
<!-- Contenedor principal --> | <!-- Contenedor principal --> | ||
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<article id="July-1" class="note-item note_active"> | <article id="July-1" class="note-item note_active"> | ||
<h2>July 1</h2><hr> | <h2>July 1</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Received and resuspended of the IDT DNA fragments</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">GS linker+αS1-casein</li> | ||
+ | <li class="list">Enterocin B</li> | ||
+ | <li class="list">Enterocin 96</li> | ||
+ | <li class="list">Bovicin HJ50</li> | ||
+ | <li class="list">Durancin TW49M</li> | ||
+ | <li class="list">Lacticin Z</li> | ||
+ | <li class="list">Leucocyclicin Q</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | <p class="note-title"></p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Received and resuspended of the IDT DNA fragments</li> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="July-2" class="note-item"> | <article id="July-2" class="note-item"> | ||
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<article id="July-3" class="note-item"> | <article id="July-3" class="note-item"> | ||
<h2>July 3</h2><hr> | <h2>July 3</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Growth curve exp.</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different temperature Condition: 37°C, 32°C, 27°C</li> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="July-4" class="note-item"> | <article id="July-4" class="note-item"> | ||
<h2>July 4</h2><hr> | <h2>July 4</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning gene of Curcumin bio-sensor</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Doing the PCR and clean-up to amplify the DNA fragment. Digesting pET30a backbone and ligating it with αS1-casein+GS linker and T7 promoter.</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">pET30a backbone</li> | ||
+ | <li class="list">DNA segment of αS1-casein+GS linker and T7 promoter.</li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="July-5" class="note-item"> | <article id="July-5" class="note-item"> | ||
<h2>July 5</h2><hr> | <h2>July 5</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning gene of Curcumin bio-sensor Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transforming the ligation product of the Curcumin bio-sensor into <i>E. coli</i> BL21 DE3 Cultivating the culture, preparing <i>E. coli</i> culture glycerol stocks, and doing the mini preparation.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">fresh colony of <i>E. coli</i> BL21 DE3 carrying the backbone pET30a containing αS1-casein + GS linker and T7 promotor gene from an overnight plate</li> |
− | + | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
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<article id="July-7" class="note-item"> | <article id="July-7" class="note-item"> | ||
<h2>July 7</h2><hr> | <h2>July 7</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression of αS1-casein</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Cultivating <i>E. coli</i> culture containing target protein αS1-casein from the <i>E. coli</i> glycerol stock and inducing it with IPTG to produce αS1-casein.</li> | ||
+ | <li class="list">Purifying the protein by His-Tag.</li> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="July-8" class="note-item"> | <article id="July-8" class="note-item"> | ||
<h2>July 8</h2><hr> | <h2>July 8</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Growth curve exp</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different pH Condition: pH = 4, 5, 6, 7, 8</li> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="July-9" class="note-item"> | <article id="July-9" class="note-item"> | ||
<h2>July 9</h2><hr> | <h2>July 9</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="July-10" class="note-item"> | <article id="July-10" class="note-item"> | ||
<h2>July 10</h2><hr> | <h2>July 10</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Chip Production</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Preparing 18 chips for bio-sensor sensitivity pretest<br> | |
− | + | 1. Dip the gold chips in 10mM Mua, RT for 4hrs.<br> | |
− | + | 2. Wash the chips with 95% EtOH three times and dry.<br> | |
− | + | 3. Add EDC+NHS mixture (100+100mM in DDW) on chips, RT for 1hrs.<br> | |
− | + | 4. DDW rinse the chips and dry.<br> | |
− | + | 5. Add αS1-casein on chips, RT for 1hrs.<br> | |
− | + | 6. Wash with PBS three times and dry.<br> | |
− | + | 6. Dip the chips in blocking solution, RT for 1.5hrs.<br> | |
− | + | 7. Wash with PBS three times and dry.</li> | |
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</ul> | </ul> | ||
</article> | </article> | ||
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<article id="July-11" class="note-item"> | <article id="July-11" class="note-item"> | ||
<h2>July 11</h2><hr> | <h2>July 11</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Bio-sensor sensitivity pretest </p> |
− | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
− | + | <li class="list">Using DPV(Differential Pulse Voltammetry) to check whether our bio-sensor can detect curcumin.<br> | |
− | + | 1. Add the diluted curcumin samples on our bio-sensor to react for 30min.<br> | |
− | + | 2. Rinse with wash buffer and dry the chips.<br> | |
− | + | 3. Wash the reference and counter electrodes with DDW, and dry them.<br> | |
− | + | 4. Set up the three electrodes system within electrocheFunctional analysisal cell.<br> | |
− | + | 5. Use the prototype of electrocheFunctional analysisal machine to measure the DPV method</li> | |
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</ul> | </ul> | ||
</article> | </article> | ||
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<article id="July-12" class="note-item"> | <article id="July-12" class="note-item"> | ||
<h2>July 12</h2><hr> | <h2>July 12</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Bio-sensor sensitivity assay –Determine Standard Curve and Create the Formula</p> |
− | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
− | + | <li class="list">Detecting the diluted standard curcumin samples by curcumin bio-sensor and made the standard curve and doing the polynomial curve fitting.</li> | |
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</ul> | </ul> | ||
</article> | </article> | ||
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<article id="July-13" class="note-item"> | <article id="July-13" class="note-item"> | ||
<h2>July 13</h2><hr> | <h2>July 13</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Bio-sensor sensitivity assay -Detecting the real Samples from Turmeric Root</p> |
− | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
− | + | <li class="list">Milling the turmeric root and dividing the powder into two groups. One of them was added with extraction buffer but not underwent the extraction protocol, and the other was added with extraction buffer but underwent the extraction process</li> | |
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</ul> | </ul> | ||
</article> | </article> | ||
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<article id="July-14" class="note-item"> | <article id="July-14" class="note-item"> | ||
<h2>July 14</h2><hr> | <h2>July 14</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="July-15" class="note-item"> | <article id="July-15" class="note-item"> | ||
<h2>July 15</h2><hr> | <h2>July 15</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Preparing competent cell <i>E. coli</i> ER2566</li> | |
− | + | </ul> | |
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− | <p class="note-title"> | + | <p class="note-title">Growth curve exp</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Observe the growth population of <i>Bacillus subtilis</i> in different salinity condition: 0.17M, 0.25M, 0.5M, 0.75M, 1.0M</li> |
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</ul> | </ul> | ||
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</article> | </article> | ||
<article id="July-16" class="note-item"> | <article id="July-16" class="note-item"> | ||
<h2>July 16</h2><hr> | <h2>July 16</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Amplify the insert gene (Pfu PCR)</li> |
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Lacticin Z + pTXB1</li> |
− | <li class="list"> | + | <li class="list">Bovincin HJ50+ pTXB1</li> |
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</ul> | </ul> | ||
− | + | </ul> | |
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</article> | </article> | ||
<article id="July-17" class="note-item"> | <article id="July-17" class="note-item"> | ||
<h2>July 17</h2><hr> | <h2>July 17</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li> |
+ | <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Lacticin Z + pTXB1</li> |
− | <li class="list"> | + | <li class="list">Bovincin HJ50+ pTXB1</li> |
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</ul> | </ul> | ||
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</article> | </article> | ||
<article id="July-18" class="note-item"> | <article id="July-18" class="note-item"> | ||
<h2>July 18</h2><hr> | <h2>July 18</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li> |
+ | <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li> | ||
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Lacticin Z + pTXB1</li> |
− | <li class="list"> | + | <li class="list">Bovincin HJ50+ pTXB1</li> |
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</ul> | </ul> | ||
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</article> | </article> | ||
<article id="July-19" class="note-item"> | <article id="July-19" class="note-item"> | ||
<h2>July 19</h2><hr> | <h2>July 19</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivation</li> |
+ | <li class="list">Miniprep (Purify Plasmid)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Lacticin Z + pTXB1</li> |
− | <li class="list"> | + | <li class="list">Bovincin HJ50+ pTXB1</li> |
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</ul> | </ul> | ||
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</article> | </article> | ||
<article id="July-20" class="note-item"> | <article id="July-20" class="note-item"> | ||
<h2>July 20</h2><hr> | <h2>July 20</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Sequencing plasmid</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Lacticin Z + pTXB1</li> |
− | <li class="list"> | + | <li class="list">Bovincin HJ50+ pTXB1</li> |
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</ul> | </ul> | ||
</ul> | </ul> | ||
Line 1,385: | Line 922: | ||
<article id="July-21" class="note-item"> | <article id="July-21" class="note-item"> | ||
<h2>July 21</h2><hr> | <h2>July 21</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
− | + | </article> | |
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<article id="July-22" class="note-item"> | <article id="July-22" class="note-item"> | ||
<h2>July 22</h2><hr> | <h2>July 22</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Amplify the insert gene (Pfu PCR)</li> |
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
+ | <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li> | ||
+ | <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pTXB1</li> |
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</ul> | </ul> | ||
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</ul> | </ul> | ||
</article> | </article> | ||
Line 1,551: | Line 942: | ||
<article id="July-23" class="note-item"> | <article id="July-23" class="note-item"> | ||
<h2>July 23</h2><hr> | <h2>July 23</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
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<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list">Transformation of | + | <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li> |
+ | <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li> | ||
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pTXB1</li> |
− | + | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
Line 1,611: | Line 956: | ||
<article id="July-24" class="note-item"> | <article id="July-24" class="note-item"> | ||
<h2>July 24</h2><hr> | <h2>July 24</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivation</li> |
+ | <li class="list">Miniprep (Purify Plasmid)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pTXB1</li> |
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</ul> | </ul> | ||
</ul> | </ul> | ||
Line 1,648: | Line 969: | ||
<article id="July-25" class="note-item"> | <article id="July-25" class="note-item"> | ||
<h2>July 25</h2><hr> | <h2>July 25</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Sequencing plasmid</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pTXB1</li> |
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</ul> | </ul> | ||
</ul> | </ul> | ||
Line 1,696: | Line 981: | ||
<article id="July-26" class="note-item"> | <article id="July-26" class="note-item"> | ||
<h2>July 26</h2><hr> | <h2>July 26</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="July-27" class="note-item"> | <article id="July-27" class="note-item"> | ||
<h2>July 27</h2><hr> | <h2>July 27</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="July-28" class="note-item"> | <article id="July-28" class="note-item"> | ||
<h2>July 28</h2><hr> | <h2>July 28</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="July-29" class="note-item"> | <article id="July-29" class="note-item"> | ||
<h2>July 29</h2><hr> | <h2>July 29</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)</li> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pTXB1</li> |
− | + | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
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</article> | </article> | ||
<article id="July-30" class="note-item"> | <article id="July-30" class="note-item"> | ||
<h2>July 30</h2><hr> | <h2>July 30</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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− | + | ||
</article> | </article> | ||
<article id="July-31" class="note-item"> | <article id="July-31" class="note-item"> | ||
Line 1,819: | Line 1,018: | ||
<article id="August-1" class="note-item"> | <article id="August-1" class="note-item"> | ||
<h2>August 1</h2><hr> | <h2>August 1</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="August-2" class="note-item"> | <article id="August-2" class="note-item"> | ||
<h2>August 2</h2><hr> | <h2>August 2</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
− | + | ||
− | + | ||
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</article> | </article> | ||
<article id="August-3" class="note-item"> | <article id="August-3" class="note-item"> | ||
<h2>August 3</h2><hr> | <h2>August 3</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
− | + | ||
− | + | ||
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</article> | </article> | ||
<article id="August-4" class="note-item"> | <article id="August-4" class="note-item"> | ||
<h2>August 4</h2><hr> | <h2>August 4</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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</article> | </article> | ||
<article id="August-5" class="note-item"> | <article id="August-5" class="note-item"> | ||
<h2>August 5</h2><hr> | <h2>August 5</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)</li> |
<li class="list">sample</li> | <li class="list">sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pTXB1</li> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
Line 1,967: | Line 1,051: | ||
<article id="August-6" class="note-item"> | <article id="August-6" class="note-item"> | ||
<h2>August 6</h2><hr> | <h2>August 6</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Cultivation (Cultivation of <i>E. coli</i> ER2566 colonies for IPTG induction)</li> | ||
+ | <li class="list">IPTG Induction (To test the O.D. levels of <i>E. coli</i> ER2566 to induce)Condition: O.D. = 0.5, 1.3</li> | ||
+ | <li class="list">sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Leucocyclicin Q + pTXB1</li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="August-7" class="note-item"> | <article id="August-7" class="note-item"> | ||
<h2>August 7</h2><hr> | <h2>August 7</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">SDS-PAGE (To check the protein production after induction)</li> | ||
+ | <li class="list">sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Leucocyclicin Q + pTXB1</li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="August-8" class="note-item"> | <article id="August-8" class="note-item"> | ||
Line 1,987: | Line 1,086: | ||
<article id="August-11" class="note-item"> | <article id="August-11" class="note-item"> | ||
<h2>August 11</h2><hr> | <h2>August 11</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Amplify the insert gene (Pfu PCR)</li> | ||
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
+ | <li class="list">sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Enterocin B</li> | ||
+ | <li class="list">Enterocin 96 </li> | ||
+ | <li class="list">Durancin TW49M </li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="August-12" class="note-item"> | <article id="August-12" class="note-item"> | ||
<h2>August 12</h2><hr> | <h2>August 12</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Digestion of PCR products (Including insert and backbone pTXB1)</li> | ||
+ | <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)</li> | ||
+ | <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li> | ||
+ | <li class="list">sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Enterocin B + pTXB1</li> | ||
+ | <li class="list">Enterocin 96 + pTXB1</li> | ||
+ | <li class="list">Durancin TW-49M + pTXB1</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | <p class="note-title">Growth curve</p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Observe the growth of <i>Bacillus subtilis</i> in different temperature, pH, salinityCondition:</li> | ||
+ | <li class="list">1.Temp: 37°C , pH:7 , salinity: 0.17M</li> | ||
+ | <li class="list">2.Temp: 30°C , pH:9 , salinity: 0.5M</li> | ||
+ | <li class="list">3.Temp: 25°C , pH:5 , salinity: 0.25M</li> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="August-13" class="note-item"> | <article id="August-13" class="note-item"> | ||
<h2>August 13</h2><hr> | <h2>August 13</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li> | ||
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
+ | <li class="list">sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Enterocin B + pTXB1</li> | ||
+ | <li class="list">Enterocin 96 + pTXB1</li> | ||
+ | <li class="list">Durancin TW-49M + pTXB1</li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
Line 2,001: | Line 1,138: | ||
<article id="August-14" class="note-item"> | <article id="August-14" class="note-item"> | ||
<h2>August 14</h2><hr> | <h2>August 14</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivation</li> |
+ | <li class="list">Miniprep (Purify Plasmid)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Enterocin B + pTXB1</li> | |
− | + | <li class="list">Enterocin 96 + pTXB1</li> | |
− | + | <li class="list">Durancin TW-49M + pTXB1</li> | |
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
− | + | </ul> | |
− | + | <p class="note-title">Expression</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transformation (Transform correct plasmid into <i>E. coli</i> ER2566)to test different cultivation temperature</li> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
</article> | </article> | ||
Line 2,030: | Line 1,157: | ||
<article id="August-15" class="note-item"> | <article id="August-15" class="note-item"> | ||
<h2>August 15</h2><hr> | <h2>August 15</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Sequencing plasmid</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Enterocin B + pTXB1</li> | |
− | + | <li class="list">Enterocin 96 + pTXB1</li> | |
− | + | <li class="list">Durancin TW-49M + pTXB1</li> | |
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivation (Cultivation of <i>E. coli</i> ER2566 colonies for IPTG induction)</li> |
− | + | <li class="list">IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C</li> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | <li class="list"> | + | |
− | + | ||
− | + | ||
− | + | ||
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− | + | ||
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− | + | ||
− | + | ||
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− | + | ||
− | + | ||
− | + | ||
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− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
</article> | </article> | ||
Line 2,095: | Line 1,177: | ||
<article id="August-16" class="note-item"> | <article id="August-16" class="note-item"> | ||
<h2>August 16</h2><hr> | <h2>August 16</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">SDS-PAGE (To check the protein production after induction)Check different temperature</li> |
− | + | <li class="list">IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM</li> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | <li class="list"> | + | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
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− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
</article> | </article> | ||
Line 2,152: | Line 1,186: | ||
<article id="August-17" class="note-item"> | <article id="August-17" class="note-item"> | ||
<h2>August 17</h2><hr> | <h2>August 17</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Expression</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list">To check the protein | + | <li class="list">SDS-PAGE (To check the protein production after induction) -> Check different IPTG concentration</li> |
+ | </ul> | ||
+ | </article> | ||
+ | |||
+ | <article id="August-18" class="note-item"> | ||
+ | <h2>August 18</h2><hr> | ||
+ | <p class="note-title">Transformation</p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Transforming backbone pTXB1 into <i>E. coli</i> ER2566 to produce protein as our negative control in verifying the function of our target peptide.</li> | ||
+ | <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>E. coli</i> ER2566 to produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">backbone pTXB1 mini</li> |
− | <li class="list"> | + | <li class="list">backbone pTXB1 containing Enterocin B gene mini</li> |
− | <li class="list"> | + | <li class="list">backbone pTXB1 containing Enterocin 96 gene mini</li> |
− | <li class="list"> | + | <li class="list">backbone pTXB1 containing Leucocyclicin Q gene mini</li> |
− | + | ||
</ul> | </ul> | ||
− | + | <p class="note-title">Cultivation</p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>Bacillus subtilis</i> ER2566 at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.</li> |
+ | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 from an overnight plate</li> |
− | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate</li> | |
− | <li class="list"> | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate</li> |
− | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate</li> | |
− | <li class="list"> | + | </ul> |
− | + | <p class="note-title">IPTG Induction</p> | |
− | <li class="list"> | + | |
− | + | ||
− | </ul> | + | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 after cultivating at 37°C to produce protein as our negative control in verifying the function of our target peptide</li> |
− | <li class="list"> | + | <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1</li> |
− | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin B gene</li> | |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene</li> |
− | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene</li> | |
− | <li class="list"> | + | </ul> |
− | + | ||
− | <li class="list"> | + | |
− | + | ||
− | + | ||
− | </ul> | + | |
− | + | ||
</article> | </article> | ||
<article id="August-19" class="note-item"> | <article id="August-19" class="note-item"> | ||
<h2>August 19</h2><hr> | <h2>August 19</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Digestion of PCR products (Including insert and backbone pET30a)</li> |
− | <li class="list"> | + | <li class="list">Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)</li> |
<li class="list">Sample(8/17)</li> | <li class="list">Sample(8/17)</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Enterocin B + pET30a</li> |
− | <li class="list"> | + | <li class="list">Enterocin 96 + pET30a</li> |
− | <li class="list"> | + | <li class="list">Bovicin HJ50 + pET30a</li> |
− | + | <li class="list">Durancin TW-49M + pET30a</li> | |
− | <li class="list"> | + | <li class="list">Lacticin Z + pET30a</li> |
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pET30a</li> |
− | + | ||
− | <li class="list"> | + | |
− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | + | <p class="note-title">Sonication</p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Breaking <i>E. coli</i> ER2566 carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 induced by IPTG</li> |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | + | <p class="note-title">Protein quantification & SDS-PAGE</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 induced by IPTG</li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li> |
</ul> | </ul> | ||
</ul> | </ul> | ||
+ | </article> | ||
− | <p class="note-title"> | + | <article id="August-20" class="note-item"> |
+ | <h2>August 20</h2><hr> | ||
+ | <p class="note-title">Cloning</p> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transformation of ligation products (Transform into <i>E. coli</i> DH5α)</li> |
+ | <li class="list">Taq PCR (PCR of ligation products to amplify insert gene)</li> | ||
+ | <li class="list">Electrophoresis (To check PCR products)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Enterocin B + pET30a</li> |
− | <li class="list"> | + | <li class="list">Enterocin 96 + pET30a</li> |
− | <li class="list"> | + | <li class="list">Bovicin HJ50 + pET30a</li> |
+ | <li class="list">Durancin TW-49M + pET30a</li> | ||
+ | <li class="list">Lacticin Z + pET30a</li> | ||
+ | <li class="list">Leucocyclicin Q + pET30a</li> | ||
</ul> | </ul> | ||
− | </ul> | + | </ul> |
</article> | </article> | ||
<article id="August-21" class="note-item"> | <article id="August-21" class="note-item"> | ||
<h2>August 21</h2><hr> | <h2>August 21</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivation</li> |
+ | <li class="list">Miniprep (Purify Plasmid)</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Enterocin B + pET30a</li> |
− | <li class="list"> | + | <li class="list">Enterocin 96 + pET30a</li> |
− | <li class="list"> | + | <li class="list">Bovicin HJ50 + pET30a</li> |
− | <li class="list"> | + | <li class="list">Durancin TW-49M + pET30a</li> |
− | <li class="list"> | + | <li class="list">Lacticin Z + pET30a</li> |
− | <li class="list"> | + | <li class="list">Leucocyclicin Q + pET30a</li> |
</ul> | </ul> | ||
− | |||
− | |||
− | |||
</article> | </article> | ||
<article id="August-22" class="note-item"> | <article id="August-22" class="note-item"> | ||
<h2>August 22</h2><hr> | <h2>August 22</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cloning</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Sequencing plasmid</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Enterocin B + pET30a</li> |
− | + | <li class="list">Enterocin 96 + pET30a</li> | |
− | + | <li class="list">Bovicin HJ50 + pET30a</li> | |
− | + | <li class="list">Durancin TW-49M + pET30a</li> | |
− | + | <li class="list">Lacticin Z + pET30a</li> | |
− | + | <li class="list">Leucocyclicin Q + pET30a</li> | |
− | + | ||
− | + | ||
− | + | ||
− | <li class="list"> | + | |
− | <li class="list"> | + | |
− | + | ||
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− | <li class="list"> | + | |
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− | <li class="list"> | + | |
− | <li class="list"> | + | |
− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
Line 2,338: | Line 1,324: | ||
<article id="August-23" class="note-item"> | <article id="August-23" class="note-item"> | ||
<h2>August 23</h2><hr> | <h2>August 23</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
− | + | ||
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</article> | </article> | ||
<article id="August-24" class="note-item"> | <article id="August-24" class="note-item"> | ||
<h2>August 24</h2><hr> | <h2>August 24</h2><hr> | ||
− | + | <p class="note-title">Transformation</p> | |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>E. coli</i> ER2566 to produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">backbone pTXB1 containing Bovicin HJ50 gene mini</li> |
+ | <li class="list">backbone pTXB1 containing Durancin TW-49M gene mini</li> | ||
</ul> | </ul> | ||
− | + | </ul> | |
− | + | <p class="note-title">Cultivation</p> | |
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate</li> |
− | <li class="list"> | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate</li> |
− | + | </ul> | |
− | + | </ul> | |
− | + | <p class="note-title">IPTG Induction</p> | |
− | + | ||
− | + | ||
− | + | ||
− | </ul> | + | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.</li> | |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene</li> |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene</li> |
</ul> | </ul> | ||
− | + | </ul> | |
− | + | ||
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</article> | </article> | ||
<article id="August-25" class="note-item"> | <article id="August-25" class="note-item"> | ||
<h2>August 25</h2><hr> | <h2>August 25</h2><hr> | ||
− | + | <p class="note-title">Sonication</p> | |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Breaking <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG </li> |
+ | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG </li> | ||
</ul> | </ul> | ||
− | |||
</ul> | </ul> | ||
− | + | <p class="note-title">Protein quantification & SDS-PAGE</p> | |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
− | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG </li> |
− | + | <li class="list">Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG </li> | |
+ | </ul> | ||
</ul> | </ul> | ||
</article> | </article> | ||
Line 2,440: | Line 1,382: | ||
<article id="August-26" class="note-item"> | <article id="August-26" class="note-item"> | ||
<h2>August 26</h2><hr> | <h2>August 26</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="August-27" class="note-item"> | <article id="August-27" class="note-item"> | ||
<h2>August 27</h2><hr> | <h2>August 27</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
+ | |||
<article id="August-28" class="note-item"> | <article id="August-28" class="note-item"> | ||
<h2>August 28</h2><hr> | <h2>August 28</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="August-29" class="note-item"> | <article id="August-29" class="note-item"> | ||
<h2>August 29</h2><hr> | <h2>August 29</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="August-30" class="note-item"> | <article id="August-30" class="note-item"> | ||
<h2>August 30</h2><hr> | <h2>August 30</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="August-31" class="note-item"> | <article id="August-31" class="note-item"> | ||
<h2>August 31</h2><hr> | <h2>August 31</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
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<article id="September-1" class="note-item"> | <article id="September-1" class="note-item"> | ||
<h2>September 1</h2><hr> | <h2>September 1</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-2" class="note-item"> | <article id="September-2" class="note-item"> | ||
<h2>September 2</h2><hr> | <h2>September 2</h2><hr> | ||
− | + | <p class="note-title">None</p> | |
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</article> | </article> | ||
<article id="September-3" class="note-item"> | <article id="September-3" class="note-item"> | ||
<h2>September 3</h2><hr> | <h2>September 3</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-4" class="note-item"> | <article id="September-4" class="note-item"> | ||
<h2>September 4</h2><hr> | <h2>September 4</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-5" class="note-item"> | <article id="September-5" class="note-item"> | ||
<h2>September 5</h2><hr> | <h2>September 5</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
− | + | </article> | |
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− | + | <article id="September-6" class="note-item"> | |
+ | <h2>September 6</h2><hr> | ||
+ | <p class="note-title">Transformation</p> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Transforming backbone pTXB1 containing Lacticin Z gene into <i>E. coli</i> ER2566 to produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">backbone pTXB1 containing Lacticin Z gene mini</li> |
− | + | ||
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</ul> | </ul> | ||
− | + | </ul> | |
− | + | <p class="note-title">Cultivation</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>Bacillus subtilis</i> ER2566 at 37°C carrying the backbone pTXB1 containing Lacticin Z gene to produce our target protein as a bio-stimulator.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Fresh colony of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate</li> |
− | + | </ul> | |
− | </ul> | + | </ul> |
− | + | <p class="note-title">IPTG Induction</p> | |
− | + | ||
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− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Inducting <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating at 37°C to produce our target protein as a bio-stimulator.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene</li> |
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</ul> | </ul> | ||
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</article> | </article> | ||
<article id="September-7" class="note-item"> | <article id="September-7" class="note-item"> | ||
<h2>September 7</h2><hr> | <h2>September 7</h2><hr> | ||
− | + | <p class="note-title">Sonication</p> | |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Breaking <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating to harvest the protein by sonication.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Culture of <i>E. coli</i> ER2566 carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG </li> | |
− | + | ||
</ul> | </ul> | ||
− | |||
</ul> | </ul> | ||
− | + | <p class="note-title">Protein quantification & SDS-PAGE</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Quantificating the protein expressing the backbone pTXB1 containing Lacticin Z gene by Bradford method and running SDS-PAGE to check the protein expression.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG </li> | |
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</ul> | </ul> | ||
</ul> | </ul> | ||
Line 3,206: | Line 1,487: | ||
<article id="September-8" class="note-item"> | <article id="September-8" class="note-item"> | ||
<h2>September 8</h2><hr> | <h2>September 8</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-9" class="note-item"> | <article id="September-9" class="note-item"> | ||
<h2>September 9</h2><hr> | <h2>September 9</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
Line 3,343: | Line 1,502: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-11" class="note-item"> | <article id="September-11" class="note-item"> | ||
<h2>September 11</h2><hr> | <h2>September 11</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
<article id="September-12" class="note-item"> | <article id="September-12" class="note-item"> | ||
<h2>September 12</h2><hr> | <h2>September 12</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
Line 3,380: | Line 1,517: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-14" class="note-item"> | <article id="September-14" class="note-item"> | ||
<h2>September 14</h2><hr> | <h2>September 14</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-15" class="note-item"> | <article id="September-15" class="note-item"> | ||
<h2>September 15</h2><hr> | <h2>September 15</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-16" class="note-item"> | <article id="September-16" class="note-item"> | ||
<h2>September 16</h2><hr> | <h2>September 16</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-17" class="note-item"> | <article id="September-17" class="note-item"> | ||
<h2>September 17</h2><hr> | <h2>September 17</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
Line 3,413: | Line 1,542: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-19" class="note-item"> | <article id="September-19" class="note-item"> | ||
<h2>September 19</h2><hr> | <h2>September 19</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-20" class="note-item"> | <article id="September-20" class="note-item"> | ||
<h2>September 20</h2><hr> | <h2>September 20</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-21" class="note-item"> | <article id="September-21" class="note-item"> | ||
<h2>September 21</h2><hr> | <h2>September 21</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Functional analysis test</p> |
− | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
− | + | <li class="list">Test the function of Lacticin Z (produced by <i>E. coli</i> ER2566) by elisa reader(triplicate)</li> | |
− | + | ||
− | + | ||
− | + | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | + | <li class="list">Control:<br> | |
− | + | 1. <i>Bacillus subtilis</i> + Ampicillin<br> | |
− | + | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br> | |
− | + | 3. <i>Bacillus subtilis</i> + LB broth<br> | |
− | + | 4. LB broth for background</li> | |
− | + | <li class="list">Test:<br> | |
+ | 1. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG</li><br> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
+ | </article> | ||
+ | |||
+ | <article id="September-22" class="note-item"> | ||
+ | <h2>September 22</h2><hr> | ||
+ | <p class="note-title">None</p> | ||
</article> | </article> | ||
<article id="September-23" class="note-item"> | <article id="September-23" class="note-item"> | ||
<h2>September 23</h2><hr> | <h2>September 23</h2><hr> | ||
− | + | <p class="note-title">None</p> | |
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</article> | </article> | ||
<article id="September-24" class="note-item"> | <article id="September-24" class="note-item"> | ||
<h2>September 24</h2><hr> | <h2>September 24</h2><hr> | ||
− | + | <p class="note-title">None</p> | |
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</article> | </article> | ||
<article id="September-25" class="note-item"> | <article id="September-25" class="note-item"> | ||
<h2>September 25</h2><hr> | <h2>September 25</h2><hr> | ||
− | + | <p class="note-title">None</p> | |
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</article> | </article> | ||
Line 3,525: | Line 1,595: | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-27" class="note-item"> | <article id="September-27" class="note-item"> | ||
<h2>September 27</h2><hr> | <h2>September 27</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-28" class="note-item"> | <article id="September-28" class="note-item"> | ||
<h2>September 28</h2><hr> | <h2>September 28</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">None</p> |
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</article> | </article> | ||
+ | |||
<article id="September-29" class="note-item"> | <article id="September-29" class="note-item"> | ||
<h2>September 29</h2><hr> | <h2>September 29</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="September-30" class="note-item"> | <article id="September-30" class="note-item"> | ||
<h2>September 30</h2><hr> | <h2>September 30</h2><hr> | ||
Line 3,554: | Line 1,619: | ||
<article id="October-1" class="note-item"> | <article id="October-1" class="note-item"> | ||
− | <h2>October | + | <h2>October 1</h2><hr> |
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-2" class="note-item"> | <article id="October-2" class="note-item"> | ||
<h2>October 2</h2><hr> | <h2>October 2</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-3" class="note-item"> | <article id="October-3" class="note-item"> | ||
<h2>October 3</h2><hr> | <h2>October 3</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-4" class="note-item"> | <article id="October-4" class="note-item"> | ||
<h2>October 4</h2><hr> | <h2>October 4</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-5" class="note-item"> | <article id="October-5" class="note-item"> | ||
<h2>October 5</h2><hr> | <h2>October 5</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Transformation</p> |
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Transforming backbone pTXB1 into <i>E. coli</i> Rosetta-Gami to produce protein as our negative control in verifying the function of our target peptide.</li> |
+ | <li class="list">Transforming backbone pTXB1 containing bacteriocin gene into <i>E. coli</i> Rosetta-Gami to produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">backbone pTXB1 mini</li> |
+ | <li class="list">backbone pTXB1 containing Enterocin B gene mini</li> | ||
+ | <li class="list">backbone pTXB1 containing Enterocin 96 gene mini</li> | ||
+ | <li class="list">backbone pTXB1 containing Bovicin HJ50 gene mini</li> | ||
+ | <li class="list">backbone pTXB1 containing Durancin TW-49M gene mini</li> | ||
+ | <li class="list">backbone pTXB1 containing Lacticin Z gene mini</li> | ||
+ | <li class="list">backbone pTXB1 containing Leucocyclicin Q gene mini</li> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
Line 3,583: | Line 1,659: | ||
<article id="October-6" class="note-item"> | <article id="October-6" class="note-item"> | ||
<h2>October 6</h2><hr> | <h2>October 6</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cultivation</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.</li> | ||
+ | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 from an overnight plate</li> | ||
+ | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate</li> | ||
+ | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate</li> | ||
+ | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | <p class="note-title">IPTG Induction</p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.</li> | ||
+ | <li class="list">Inducting <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 from an overnight plate</li> | ||
+ | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene</li> | ||
+ | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene</li> | ||
+ | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene</li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
<article id="October-7" class="note-item"> | <article id="October-7" class="note-item"> | ||
<h2>October 7</h2><hr> | <h2>October 7</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Cultivation</p> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Cultivate the <i>E. coli</i> ER2566 at 37°C cultivating <i>E. coli</i> Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate</li> |
− | + | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate</li> | |
− | + | <li class="list">Fresh colony of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate</li> | |
− | <li class="list"> | + | |
− | + | ||
− | + | ||
− | <li class="list"> | + | |
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | + | <p class="note-title">IPTG Induction</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Inducting <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene</li> |
− | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene</li> | |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene</li> |
− | + | ||
− | + | ||
− | <li class="list"> | + | |
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | + | <p class="note-title">Sonication</p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Breaking <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 induced by IPTG </li> |
− | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG</li> | |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG</li> |
− | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG</li> | |
− | <li class="list"> | + | |
− | + | ||
− | <li class="list"> | + | |
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | + | <p class="note-title">Protein quantification & SDS-PAGE</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 induced by IPTG</li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li> |
− | + | ||
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
+ | </article> | ||
− | <p class="note-title"> | + | <article id="October-8" class="note-item"> |
+ | <h2>October 8</h2><hr> | ||
+ | <p class="note-title">Sonication</p> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Breaking <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG</li> |
− | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG</li> | |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG</li> |
− | <li class="list"> | + | <li class="list">Culture of <i>E. coli</i> Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG</li> |
− | + | ||
− | <li class="list"> | + | |
− | + | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
− | + | <p class="note-title">Protein quantification & SDS-PAGE</p> | |
− | + | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.</li> | ||
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG </li> |
− | <li class="list"> | + | <li class="list">Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG </li> |
− | + | </ul> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</ul> | </ul> | ||
+ | </article> | ||
+ | |||
+ | <article id="October-9" class="note-item"> | ||
+ | <h2>October 9</h2><hr> | ||
+ | <p class="note-title">Functional analysis test</p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(triplicate)</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Control:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Ampicillin<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br> | ||
+ | 3. <i>Bacillus subtilis</i> + LB broth<br> | ||
+ | 4. LB broth for background</li> | ||
+ | <li class="list">Test:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG<br> | ||
+ | 3. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG<br> | ||
+ | 4. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | </article> | ||
+ | |||
+ | <article id="October-10" class="note-item"> | ||
+ | <h2>October 10</h2><hr> | ||
+ | <p class="note-title">Functional analysis test</p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(triplicate)</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Control:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Ampicillin<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br> | ||
+ | 3. <i>Bacillus subtilis</i> + LB broth<br> | ||
+ | 4. LB broth for background</li> | ||
+ | <li class="list">Test:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | </article> | ||
+ | |||
+ | <article id="October-11" class="note-item"> | ||
+ | <h2>October 11</h2><hr> | ||
+ | <p class="note-title">None</p> | ||
</article> | </article> | ||
<article id="October-12" class="note-item"> | <article id="October-12" class="note-item"> | ||
<h2>October 12</h2><hr> | <h2>October 12</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Functional analysis test</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(Dose response assessment)</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Control:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Ampicillin<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br> | ||
+ | 3. <i>Bacillus subtilis</i> + LB broth<br> | ||
+ | 4. LB broth for background</li> | ||
+ | <li class="list">Test:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG<br> | ||
+ | 3. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG<br> | ||
+ | 4. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG</li> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
+ | |||
<article id="October-13" class="note-item"> | <article id="October-13" class="note-item"> | ||
<h2>October 13</h2><hr> | <h2>October 13</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-14" class="note-item"> | <article id="October-14" class="note-item"> | ||
<h2>October 14</h2><hr> | <h2>October 14</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-15" class="note-item"> | <article id="October-15" class="note-item"> | ||
<h2>October 15</h2><hr> | <h2>October 15</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Functional analysis test</p> |
− | + | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | |
− | <li class="list"> | + | <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(Dose response assessment)</li> |
<li class="list">Sample</li> | <li class="list">Sample</li> | ||
<ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
− | <li class="list"> | + | <li class="list">Control:<br> |
− | <li class="list"> | + | 1. <i>Bacillus subtilis</i> + Ampicillin<br> |
− | + | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br> | |
− | + | 3. <i>Bacillus subtilis</i> + LB broth<br> | |
− | <li class="list"> | + | 4. LB broth for background</li> |
− | <li class="list"> | + | <li class="list">Test:<br> |
+ | 1. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG</li> | ||
+ | </ul> | ||
+ | </ul> | ||
+ | <p class="note-title">Functional analysis test(inhibition zone)</p> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Test the function of Enterocin B (produced by <i>E. coli</i> Rosetta-gami) by LB Agar + Ampicillin plate</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">The <i>Bacillus subtilis</i> on the plate</li> | ||
+ | <li class="list">Control:<br> | ||
+ | 1. Ampicillin<br> | ||
+ | 2. Production of the pTXB1 backbone induced by IPTG<br> | ||
+ | <li class="list">Test:<br> | ||
+ | 1. Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG<br> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
Line 3,729: | Line 1,866: | ||
<article id="October-16" class="note-item"> | <article id="October-16" class="note-item"> | ||
<h2>October 16</h2><hr> | <h2>October 16</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Functional analysis test(inhibition zone)</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by LB Agar + Ampicillin plate</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">The <i>Bacillus subtilis</i> on the plate</li> | ||
+ | <li class="list">Control:<br> | ||
+ | 1. Ampicillin<br> | ||
+ | 2. Production of the pTXB1 backbone induced by IPTG<br> | ||
+ | <li class="list">Test:<br> | ||
+ | 1. Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG<br> | ||
+ | 2. Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG<br> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
+ | |||
<article id="October-17" class="note-item"> | <article id="October-17" class="note-item"> | ||
<h2>October 17</h2><hr> | <h2>October 17</h2><hr> | ||
− | <p class="note-title"> | + | <p class="note-title">Functional analysis test</p> |
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Test the function of bacteriocins (produced by <i>E. coli</i> Rosetta-gami) by elisa reader(Dose response assessment)</li> | ||
+ | <li class="list">Sample</li> | ||
+ | <ul style="list-style-image:none;list-style-type: disc;padding-left:12px;"> | ||
+ | <li class="list">Control:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Ampicillin<br> | ||
+ | 2. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone induced by IPTG<br> | ||
+ | 3. <i>Bacillus subtilis</i> + LB broth<br> | ||
+ | <li class="list">Test:<br> | ||
+ | 1. <i>Bacillus subtilis</i> + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG<br> | ||
+ | </ul> | ||
+ | </ul> | ||
</article> | </article> | ||
+ | |||
<article id="October-18" class="note-item"> | <article id="October-18" class="note-item"> | ||
<h2>October 18</h2><hr> | <h2>October 18</h2><hr> | ||
<p class="note-title">None</p> | <p class="note-title">None</p> | ||
</article> | </article> | ||
+ | |||
<article id="October-19" class="note-item"> | <article id="October-19" class="note-item"> | ||
<h2>October 19</h2><hr> | <h2>October 19</h2><hr> |
Latest revision as of 19:43, 17 October 2018
Wet Lab
July 1
Cloning
- Received and resuspended of the IDT DNA fragments
- Sample
- GS linker+αS1-casein
- Enterocin B
- Enterocin 96
- Bovicin HJ50
- Durancin TW49M
- Lacticin Z
- Leucocyclicin Q
- Received and resuspended of the IDT DNA fragments
July 2
None
July 3
Growth curve exp.
- Observe the growth population of Bacillus subtilis in different temperature Condition: 37°C, 32°C, 27°C
July 4
Cloning gene of Curcumin bio-sensor
- Doing the PCR and clean-up to amplify the DNA fragment. Digesting pET30a backbone and ligating it with αS1-casein+GS linker and T7 promoter.
- Sample
- pET30a backbone
- DNA segment of αS1-casein+GS linker and T7 promoter.
July 5
Cloning gene of Curcumin bio-sensor Expression
- Transforming the ligation product of the Curcumin bio-sensor into E. coli BL21 DE3 Cultivating the culture, preparing E. coli culture glycerol stocks, and doing the mini preparation.
- Sample
- fresh colony of E. coli BL21 DE3 carrying the backbone pET30a containing αS1-casein + GS linker and T7 promotor gene from an overnight plate
July 6
None
July 7
Expression of αS1-casein
- Cultivating E. coli culture containing target protein αS1-casein from the E. coli glycerol stock and inducing it with IPTG to produce αS1-casein.
- Purifying the protein by His-Tag.
July 8
Growth curve exp
- Observe the growth population of Bacillus subtilis in different pH Condition: pH = 4, 5, 6, 7, 8
July 9
None
July 10
Chip Production
- Preparing 18 chips for bio-sensor sensitivity pretest
1. Dip the gold chips in 10mM Mua, RT for 4hrs.
2. Wash the chips with 95% EtOH three times and dry.
3. Add EDC+NHS mixture (100+100mM in DDW) on chips, RT for 1hrs.
4. DDW rinse the chips and dry.
5. Add αS1-casein on chips, RT for 1hrs.
6. Wash with PBS three times and dry.
6. Dip the chips in blocking solution, RT for 1.5hrs.
7. Wash with PBS three times and dry.
July 11
Bio-sensor sensitivity pretest
- Using DPV(Differential Pulse Voltammetry) to check whether our bio-sensor can detect curcumin.
1. Add the diluted curcumin samples on our bio-sensor to react for 30min.
2. Rinse with wash buffer and dry the chips.
3. Wash the reference and counter electrodes with DDW, and dry them.
4. Set up the three electrodes system within electrocheFunctional analysisal cell.
5. Use the prototype of electrocheFunctional analysisal machine to measure the DPV method
July 12
Bio-sensor sensitivity assay –Determine Standard Curve and Create the Formula
- Detecting the diluted standard curcumin samples by curcumin bio-sensor and made the standard curve and doing the polynomial curve fitting.
July 13
Bio-sensor sensitivity assay -Detecting the real Samples from Turmeric Root
- Milling the turmeric root and dividing the powder into two groups. One of them was added with extraction buffer but not underwent the extraction protocol, and the other was added with extraction buffer but underwent the extraction process
July 14
None
July 15
Expression
- Preparing competent cell E. coli ER2566
Growth curve exp
- Observe the growth population of Bacillus subtilis in different salinity condition: 0.17M, 0.25M, 0.5M, 0.75M, 1.0M
July 16
Cloning
- Amplify the insert gene (Pfu PCR)
- Electrophoresis (To check PCR products)
- Sample
- Lacticin Z + pTXB1
- Bovincin HJ50+ pTXB1
July 17
Cloning
- Digestion of PCR products (Including insert and backbone pTXB1)
- Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
- Sample
- Lacticin Z + pTXB1
- Bovincin HJ50+ pTXB1
July 18
Cloning
- Transformation of ligation products (Transform into E. coli DH5α)
- Taq PCR (PCR of ligation products to amplify insert gene)
- Electrophoresis (To check PCR products)
- Sample
- Lacticin Z + pTXB1
- Bovincin HJ50+ pTXB1
July 19
Cloning
- Cultivation
- Miniprep (Purify Plasmid)
- Sample
- Lacticin Z + pTXB1
- Bovincin HJ50+ pTXB1
July 20
Cloning
- Sequencing plasmid
- Sample
- Lacticin Z + pTXB1
- Bovincin HJ50+ pTXB1
July 21
None
July 22
Cloning
- Amplify the insert gene (Pfu PCR)
- Electrophoresis (To check PCR products)
- Digestion of PCR products (Including insert and backbone pTXB1)
- Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
- Sample
- Leucocyclicin Q + pTXB1
July 23
Cloning
- Transformation of ligation products (Transform into E. coli DH5α)
- Taq PCR (PCR of ligation products to amplify insert gene)
- Electrophoresis (To check PCR products)
- Sample
- Leucocyclicin Q + pTXB1
July 24
Cloning
- Cultivation
- Miniprep (Purify Plasmid)
- Sample
- Leucocyclicin Q + pTXB1
July 25
Cloning
- Sequencing plasmid
- Sample
- Leucocyclicin Q + pTXB1
July 26
None
July 27
None
July 28
None
July 29
Expression
- Transformation (Transform correct plasmid into E. coli ER2566)
- Leucocyclicin Q + pTXB1
July 30
None
July 31
None
August 1
None
August 2
None
August 3
None
August 4
None
August 5
Expression
- Transformation (Transform correct plasmid into E. coli ER2566)
- sample
- Leucocyclicin Q + pTXB1
August 6
Expression
- Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)
- IPTG Induction (To test the O.D. levels of E. coli ER2566 to induce)Condition: O.D. = 0.5, 1.3
- sample
- Leucocyclicin Q + pTXB1
August 7
Expression
- SDS-PAGE (To check the protein production after induction)
- sample
- Leucocyclicin Q + pTXB1
August 8
None
August 9
None
August 10
None
August 11
Cloning
- Amplify the insert gene (Pfu PCR)
- Electrophoresis (To check PCR products)
- sample
- Enterocin B
- Enterocin 96
- Durancin TW49M
August 12
Cloning
- Digestion of PCR products (Including insert and backbone pTXB1)
- Ligation of digestion products (Ligase the DNA fragment into backbone pTXB1)
- Transformation of ligation products (Transform into E. coli DH5α)
- sample
- Enterocin B + pTXB1
- Enterocin 96 + pTXB1
- Durancin TW-49M + pTXB1
Growth curve
- Observe the growth of Bacillus subtilis in different temperature, pH, salinityCondition:
- 1.Temp: 37°C , pH:7 , salinity: 0.17M
- 2.Temp: 30°C , pH:9 , salinity: 0.5M
- 3.Temp: 25°C , pH:5 , salinity: 0.25M
August 13
Cloning
- Taq PCR (PCR of ligation products to amplify insert gene)
- Electrophoresis (To check PCR products)
- sample
- Enterocin B + pTXB1
- Enterocin 96 + pTXB1
- Durancin TW-49M + pTXB1
August 14
Cloning
- Cultivation
- Miniprep (Purify Plasmid)
- Sample
- Enterocin B + pTXB1
- Enterocin 96 + pTXB1
- Durancin TW-49M + pTXB1
Expression
- Transformation (Transform correct plasmid into E. coli ER2566)to test different cultivation temperature
August 15
Cloning
- Sequencing plasmid
- Sample
- Enterocin B + pTXB1
- Enterocin 96 + pTXB1
- Durancin TW-49M + pTXB1
Expression
- Cultivation (Cultivation of E. coli ER2566 colonies for IPTG induction)
- IPTG Induction (To test the temperature of cultivation after induction)Condition: temp = 37, 30, 13.5°C
August 16
Expression
- SDS-PAGE (To check the protein production after induction)Check different temperature
- IPTG Induction (To test the concentration of IPTG)Condition: concentration = 200, 400, 600, 800, 1000μM
August 17
Expression
- SDS-PAGE (To check the protein production after induction) -> Check different IPTG concentration
August 18
Transformation
- Transforming backbone pTXB1 into E. coli ER2566 to produce protein as our negative control in verifying the function of our target peptide.
- Transforming backbone pTXB1 containing bacteriocin gene into E. coli ER2566 to produce our target protein as a bio-stimulator.
- Sample
- backbone pTXB1 mini
- backbone pTXB1 containing Enterocin B gene mini
- backbone pTXB1 containing Enterocin 96 gene mini
- backbone pTXB1 containing Leucocyclicin Q gene mini
- Cultivate the E. coli ER2566 at 37°C cultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.
- Cultivate the E. coli ER2566 at 37°C cultivating E. coli ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.
- Sample
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 from an overnight plate
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate
- Inducting E. coli ER2566 carrying the backbone pTXB1 after cultivating at 37°C to produce protein as our negative control in verifying the function of our target peptide
- Inducting E. coli ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.
- Sample
- Culture of E. coli ER2566 carrying the backbone pTXB1
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin B gene
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene
Cultivation
IPTG Induction
August 19
Cloning
- Digestion of PCR products (Including insert and backbone pET30a)
- Ligation of digestion products (Ligase the DNA fragment into backbone pET30a)
- Sample(8/17)
- Enterocin B + pET30a
- Enterocin 96 + pET30a
- Bovicin HJ50 + pET30a
- Durancin TW-49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
Sonication
- Breaking E. coli ER2566 carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of E. coli ER2566 carrying the backbone pTXB1 induced by IPTG
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG
Protein quantification & SDS-PAGE
- Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
- Sample
- Protein expressing the backbone pTXB1 induced by IPTG
- Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG
August 20
Cloning
- Transformation of ligation products (Transform into E. coli DH5α)
- Taq PCR (PCR of ligation products to amplify insert gene)
- Electrophoresis (To check PCR products)
- Sample
- Enterocin B + pET30a
- Enterocin 96 + pET30a
- Bovicin HJ50 + pET30a
- Durancin TW-49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
August 21
Cloning
- Cultivation
- Miniprep (Purify Plasmid)
- Sample
- Enterocin B + pET30a
- Enterocin 96 + pET30a
- Bovicin HJ50 + pET30a
- Durancin TW-49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
August 22
Cloning
- Sequencing plasmid
- Sample
- Enterocin B + pET30a
- Enterocin 96 + pET30a
- Bovicin HJ50 + pET30a
- Durancin TW-49M + pET30a
- Lacticin Z + pET30a
- Leucocyclicin Q + pET30a
August 23
None
August 24
Transformation
- Transforming backbone pTXB1 containing bacteriocin gene into E. coli ER2566 to produce our target protein as a bio-stimulator.
- Sample
- backbone pTXB1 containing Bovicin HJ50 gene mini
- backbone pTXB1 containing Durancin TW-49M gene mini
Cultivation
- Cultivate the E. coli ER2566 at 37°C cultivating E. coli ER2566 at 37°C carrying the backbone pTXB1 containing bacteriocin gene to produce our target protein as a bio-stimulator.
- Sample
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate
IPTG Induction
- Inducting E. coli ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C to produce our target protein as a bio-stimulator.
- Sample
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene
August 25
Sonication
- Breaking E. coli ER2566 carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG
Protein quantification & SDS-PAGE
- Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
- Sample
- Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG
August 26
None
August 27
None
August 28
None
August 29
None
August 30
None
August 31
None
September 1
None
September 2
None
September 3
None
September 4
None
September 5
None
September 6
Transformation
- Transforming backbone pTXB1 containing Lacticin Z gene into E. coli ER2566 to produce our target protein as a bio-stimulator.
- Sample
- backbone pTXB1 containing Lacticin Z gene mini
Cultivation
- Cultivate the E. coli ER2566 at 37°C cultivating Bacillus subtilis ER2566 at 37°C carrying the backbone pTXB1 containing Lacticin Z gene to produce our target protein as a bio-stimulator.
- Sample
- Fresh colony of E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate
IPTG Induction
- Inducting E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating at 37°C to produce our target protein as a bio-stimulator.
- Sample
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene
September 7
Sonication
- Breaking E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of E. coli ER2566 carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG
Protein quantification & SDS-PAGE
- Quantificating the protein expressing the backbone pTXB1 containing Lacticin Z gene by Bradford method and running SDS-PAGE to check the protein expression.
- Sample
- Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG
September 8
None
September 9
None
September 10
None
September 11
None
September 12
None
September 13
None
September 14
None
September 15
None
September 16
None
September 17
None
September 18
None
September 19
None
September 20
None
September 21
Functional analysis test
- Test the function of Lacticin Z (produced by E. coli ER2566) by elisa reader(triplicate)
- Sample
- Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background - Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG
September 22
None
September 23
None
September 24
None
September 25
None
September 26
None
September 27
None
September 28
None
September 29
None
September 30
None
October 1
None
October 2
None
October 3
None
October 4
None
October 5
Transformation
- Transforming backbone pTXB1 into E. coli Rosetta-Gami to produce protein as our negative control in verifying the function of our target peptide.
- Transforming backbone pTXB1 containing bacteriocin gene into E. coli Rosetta-Gami to produce our target protein as a bio-stimulator.
- Sample
- backbone pTXB1 mini
- backbone pTXB1 containing Enterocin B gene mini
- backbone pTXB1 containing Enterocin 96 gene mini
- backbone pTXB1 containing Bovicin HJ50 gene mini
- backbone pTXB1 containing Durancin TW-49M gene mini
- backbone pTXB1 containing Lacticin Z gene mini
- backbone pTXB1 containing Leucocyclicin Q gene mini
October 6
Cultivation
- Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.
- Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.
- Sample
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 from an overnight plate
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene from an overnight plate
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene from an overnight plate
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene from an overnight plate
IPTG Induction
- Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 to produce protein as our negative control in verifying the function of our target peptide.
- Inducting E. coli Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.
- Sample
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 from an overnight plate
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene
October 7
Cultivation
- Cultivate the E. coli ER2566 at 37°C cultivating E. coli Rosetta-Gami at 37°C carrying the backbone pTXB1 containing bacteriocin gene produce our target protein as a bio-stimulator.
- Sample
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene from an overnight plate
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene from an overnight plate
- Fresh colony of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene from an overnight plate
IPTG Induction
- Inducting E. coli Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating at 37°C produce our target protein as a bio-stimulator.
- Sample
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene
Sonication
- Breaking E. coli Rosetta-Gami carrying the backbone pTXB1 and pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 induced by IPTG
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin B gene induced by IPTG
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG
Protein quantification & SDS-PAGE
- Quantificating the protein expressing the backbone pTXB1 and pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
- Sample
- Protein expressing the backbone pTXB1 induced by IPTG
- Protein expressing the backbone pTXB1 containing Enterocin B gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Enterocin 96 gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG
October 8
Sonication
- Breaking E. coli Rosetta-Gami carrying the backbone pTXB1 containing bacteriocin gene after cultivating to harvest the protein by sonication.
- Sample
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Lacticin Z gene induced by IPTG
- Culture of E. coli Rosetta-Gami carrying the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG
Protein quantification & SDS-PAGE
- Quantificating the protein expressing the backbone pTXB1 containing bacteriocin gene by Bradford method and running SDS-PAGE to check the protein expression.
- Sample
- Protein expressing the backbone pTXB1 containing Bovicin HJ50 gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Durancin TW-49M gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Lacticin Z gene induced by IPTG
- Protein expressing the backbone pTXB1 containing Leucocyclicin Q gene induced by IPTG
October 9
Functional analysis test
- Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(triplicate)
- Sample
- Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background - Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG
3. Bacillus subtilis + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG
4. Bacillus subtilis + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG
October 10
Functional analysis test
- Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(triplicate)
- Sample
- Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background - Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG
October 11
None
October 12
Functional analysis test
- Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(Dose response assessment)
- Sample
- Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background - Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Enterocin 96 gene induced by IPTG
3. Bacillus subtilis + Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG
4. Bacillus subtilis + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG
October 13
None
October 14
None
October 15
Functional analysis test
- Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(Dose response assessment)
- Sample
- Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
4. LB broth for background - Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG
2. Bacillus subtilis + Production of the pTXB1 backbone containing Leucocyclicin Q gene induced by IPTG
Functional analysis test(inhibition zone)
- Test the function of Enterocin B (produced by E. coli Rosetta-gami) by LB Agar + Ampicillin plate
- Sample
- The Bacillus subtilis on the plate
- Control:
1. Ampicillin
2. Production of the pTXB1 backbone induced by IPTG
- Test:
1. Production of the pTXB1 backbone containing Enterocin B gene induced by IPTG
October 16
Functional analysis test(inhibition zone)
- Test the function of bacteriocins (produced by E. coli Rosetta-gami) by LB Agar + Ampicillin plate
- Sample
- The Bacillus subtilis on the plate
- Control:
1. Ampicillin
2. Production of the pTXB1 backbone induced by IPTG
- Test:
1. Production of the pTXB1 backbone containing Bovicin HJ50 gene induced by IPTG
2. Production of the pTXB1 backbone containing Lacticin Z gene induced by IPTG
October 17
Functional analysis test
- Test the function of bacteriocins (produced by E. coli Rosetta-gami) by elisa reader(Dose response assessment)
- Sample
- Control:
1. Bacillus subtilis + Ampicillin
2. Bacillus subtilis + Production of the pTXB1 backbone induced by IPTG
3. Bacillus subtilis + LB broth
- Test:
1. Bacillus subtilis + Production of the pTXB1 backbone containing Durancin TW-49M gene induced by IPTG
October 18
None
October 19
None