Difference between revisions of "Team:CCU Taiwan/Notebook"

 
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<div class="inner">
 
<div class="inner">
 
<header>
 
<header>
−
<h1><a href="https://2017.igem.org/Team:CCU_Taiwan" id="logo">Attributions</a></h1>
+
<h1><a href="https://2017.igem.org/Team:CCU_Taiwan" id="logo">Notebook</a></h1>
  
  
Line 108: Line 108:
 
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Demonstrate">Demonstrate</a></li>
 
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Demonstrate">Demonstrate</a></li>
 
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Applied_Design">Applied design</a></li>
 
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Applied_Design">Applied design</a></li>
 +
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/InterLab">InterLab</a></li>
 +
 
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Notebook">Notebook</a></li>
 
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Notebook">Notebook</a></li>
  
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</li>
 
</li>
  
−
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/InterLab">InterLab</a></li>
+
<li><a href="https://2017.igem.org/Team:CCU_Taiwan/Medals">Medals</a></li>
 
 
 
</ul>
 
</ul>
Line 186: Line 188:
  
 
<div class="div_nav">
 
<div class="div_nav">
−
<nav class="toc_nav" id="toc_show">
+
<nav style="top: 16vh !important;" class="toc_nav" id="toc_show">
 
<ul>
 
<ul>
 
<li>
 
<li>
−
<a href="#Fluorescein">Fluorescein curve</a>
+
<a href="#Week1">Week 01</a>
−
<ul>
+
−
<li><a href="#Fluorescein-Plate-reader">Plate reader</a></li>
+
−
<li><a href="#Fluorescein-Material">Material</a></li>
+
−
<li><a href="#Fluorescein-Method">Method</a></li>
+
−
<li><a href="#Fluorescein-Data-result">Data result</a></li>
+
−
</ul>
+
 
</li>
 
</li>
−
<li>
+
−
<a href="#OD600">OD600 Reference point</a>
+
<li>
−
      <ul>
+
<a href="#Week2">Week 02</a>
−
<li><a href="#OD600-Plate-reader">Plate reader</a></li>
+
−
<li><a href="#OD600-Material">Material</a></li>
+
−
<li><a href="#OD600-Method">Method</a></li>
+
−
<li><a href="#OD600-Data-result">Data result</a></li>
+
−
      </ul>
+
 
</li>
 
</li>
−
 
+
<li>
−
<li>
+
<a href="#Week3">Week 03</a>
−
<a href="#Cell">Cell measure</a>
+
−
      <ul>
+
</li>
−
<li><a href="#Cell-Material">Material</a></li>
+
<li>
−
<li><a href="#Cell-Method">Method</a></li>
+
<a href="#Week4">Week 04</a>
−
<li><a href="#Cell-Data-result">Data result</a></li>
+
−
      </ul>
+
</li>
 +
<li>
 +
<a href="#Week5">Week 05</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week6">Week 06</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week7">Week 07</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week8">Week 08</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week9">Week 09</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week10">Week 10</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week11">Week 11</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week12">Week 12</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week13">Week 13</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week14">Week 14</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week15">Week 15</a>
 +
 +
</li>
 +
<li>
 +
<a href="#Week16">Week 16</a>
 +
 
</li>
 
</li>
−
 
  
 
</ul>
 
</ul>
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<article class="contents_nav">
 
<article class="contents_nav">
 
   <section>
 
   <section>
−
 
+
<div id="Week1">
−
 
+
−
<div id="Fluorescein">
+
−
<h2>Fluorescein Fluorescence standard curve</h2>
+
−
 
+
−
</div>
+
−
 
+
−
<div id="Fluorescein-Plate-reader">
+
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
<h3>Plate reader</h3>
+
<h3>Week 01 (2017/07/02~2017/07/08)</h3>
−
 
+
<ol><li>Lactate detection system</li></ol>
−
    <p>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of lactate detector</p>
−
      microplate reader FLUOstar Omega</br>
+
<ol><li>CSP detection system</li></ol>
−
emission filter: 520 nm</br>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of CSP detector</p>
−
excitation filter: 485 nm
+
−
    </p>
+
−
 
+
 
</div>
 
</div>
−
 
 
−
<div id="Fluorescein-Material">
 
−
<div class="aaa"></div>
 
−
<h3>Material</h3>
 
−
<p>
 
−
Fluorescein sodium salt</br>
 
−
1xPBS</br>
 
−
Tissue culture testplate (black with flat bottom)
 
−
</p>
 
  
−
</div>
+
<div id="Week2">
−
 
+
−
<div id="Fluorescein-Method">
+
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
                <h3>Method</h3>
+
<h3>Week 02 (2017/07/09~2017/07/15)</h3>
−
+
<ol><li>Lactate detection system</li></ol>
−
<ol><li>Prepare fluorescein stock solution</li></ol>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of lactate detector</p>
 +
<ol><li>CSP detection system</li></ol>
 
<p>
 
<p>
−
1. Spin down fluorescein stock tube to make sure pellet is at the bottom of tube.</br>  
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of CSP detector<br/>
−
2. Prepare 2x fluorescein stock solution (100 μM) by resuspending fluorescein in 1 mL of 1xPBS. </br>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Request 2 backbone plasmids of <i>Bacillus subtilis</i> from iGEM
−
3. Dilute the 2x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution and resulting concentration of fluorescein  stock solution 50 μM </br></br>
+
 
</p>
 
</p>
−
<ol><li>Serial dilutions</li></ol>
+
<style type="text/css">
−
<p>
+
.tg  {border-collapse:collapse;border-spacing:0;border-color:#ccc;}
−
1. Add 100 μl of PBS into wells A2, B2, C2, D2....A12, B12, C12, D12</br>
+
.tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;border-color:#ccc;color:#333;background-color:#fff;}
−
2. Add 200 μl of fluorescein 1x stock solution into A1, B1, C1, D1</br>
+
.tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;border-color:#ccc;color:#333;background-color:#f0f0f0;}
−
3. Transfer 100 μl of fluorescein stock solution from A1 into A2. </br>
+
.tg .tg-yw4l{vertical-align:top}
−
4. Mix A2 by pipetting up and down 3x and transfer 100 μl into A3. </br>
+
.tg .tg-b7b8{background-color:#f9f9f9;vertical-align:top}
−
5. Mix A3 by pipetting up and down 3x and transfer 100 μl into A4. </br>
+
</style>
−
6. Mix A4 by pipetting up and down 3x and transfer 100 μl into A5. </br>
+
<table class="tg">
−
7. Mix A5 by pipetting up and down 3x and transfer 100 μl into A6. </br>
+
  <tr>
−
8. Mix A6 by pipetting up and down 3x and transfer 100 μl into A7. </br>
+
    <th class="tg-yw4l">Part requesting #1</th>
−
9. Mix A7 by pipetting up and down 3x and transfer 100 μl into A8. </br>
+
    <th class="tg-yw4l">BBa_K823022</th>
−
10. Mix A8 by pipetting up and down 3x and transfer 100 μl into A9. </br>
+
    <th class="tg-yw4l">pSBBs4S</th>
−
11. Mix A9 by pipetting up and down 3x and transfer 100 μl into A10.</br>  
+
  </tr>
−
12. Mix A10 by pipetting up and down 3x and transfer 100 μl into A11. </br>
+
  <tr>
−
13. Mix A11 by pipetting up and down 3x and transfer 100 μl into liquid waste.
+
    <td class="tg-b7b8">Part requesting #2</td>
−
  (Caution: Do not to continue serial dilution into column 12.)</br>
+
    <td class="tg-b7b8">BBa_K823023</td>
−
</p>
+
    <td class="tg-b7b8">pSBBs1C</td>
−
<ol><li>repeat serial dilute for Row B、D、E</strong></li></ol>
+
  </tr>
−
<ol><li>Measure fluorescence of all samples in all standard measurement modes in instrument Record the data in your notebook</strong></li></ol>
+
</table>
−
<ol><li>Import data into Excel (fluorescein standard curve tab ) Sheet_1 provided</li></ol>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Order chemical materials for preparing competent cell of <i>Bacillus subtilis</i></p>
−
+
</div>
−
<br/><br/>
+
  
 +
<div id="Week3">
 +
<div class="aaa"></div>
 +
<h3>Week 03 (2017/07/16~2017/07/22)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of lactate detector</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of CSP detector</p>
 
</div>
 
</div>
  
−
<div id="Fluorescein-Data-result">
+
<div id="Week4">
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
<h3>Data result</h3>
+
<h3>Week 04 (2017/07/23~2017/07/29)</h3>
−
<br/>
+
<ol><li>Lactate detection system</li></ol>
−
<img src="https://static.igem.org/mediawiki/2017/8/85/FFs_1.png" style="display:block; margin:auto;"><br/><br/>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Send orders to IDT for DNA synthesis</p>
−
<img src="https://static.igem.org/mediawiki/2017/a/a4/FFs_2.png" style="display:block; margin:auto;"><br/><br/>
+
<ol><li>CSP detection system</li></ol>
−
<img src="https://static.igem.org/mediawiki/2017/5/5c/FFs_3.png" style="display:block; margin:auto;"><br/>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Gene design of CSP detector</p>
 
</div>
 
</div>
−
</section>
 
  
−
<section>
 
  
−
 
+
<div id="Week5">
−
<div id="OD600">
+
<div class="aaa"></div>
−
<h2>OD600 Reference point</h2>
+
<h3>Week 05 (2017/07/30~2017/08/05)</h3>
 +
<ol><li>Participate the 5th iGEM Asia-Pacific Conference at NCTU</li></ol>
  
 
</div>
 
</div>
  
  
−
<div id="OD600-Plate-reader">
+
<div id="Week6">
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
<h3>Plate reader</h3>
+
<h3>Week 06 (2017/08/06~2017/08/12)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Prepare competent cell of <i>Bacillus subtilis</i><br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;- Amplify two backbone plasmids of <i>Bacillus subtilis</i>
 +
</p>
 +
<ol><li>InterLab Study</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Transform eight kinds of plasmids into <i>E. coli DH5α</i></p>
 +
<style type="text/css">
 +
.tg  {border-collapse:collapse;border-spacing:0;}
 +
.tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
.tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
.tg .tg-yw4l{vertical-align:top}
 +
</style>
 +
<table class="tg">
 +
  <tr>
 +
    <th class="tg-031e">Positive Control</th>
 +
    <th class="tg-031e">BBa_I20270</th>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-031e">Negative Control</td>
 +
    <td class="tg-031e">BBa_R0040</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-yw4l">Test Device 1</td>
 +
    <td class="tg-yw4l">BBa_J364000</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-yw4l">Test Device 2</td>
 +
    <td class="tg-yw4l">BBa_J364001</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-yw4l">Test Device 3</td>
 +
    <td class="tg-yw4l">BBa_J364002</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-yw4l">Test Device 4</td>
 +
    <td class="tg-yw4l">BBa_J364003</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-yw4l">Test Device 5</td>
 +
    <td class="tg-yw4l">BBa_J364004</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-yw4l">Test Device 6</td>
 +
    <td class="tg-yw4l">BBa_J364005</td>
 +
  </tr>
 +
</table>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Plate culture eight kinds of <i>E. coli DH5α</i></p>
 +
</div>
  
−
    <p>
 
−
Thermo Scientific™ Multiskan™ FC Filter-based Microplate Photometer</br>
 
−
Filter: 595 nm</br>
 
−
    </p>
 
  
 +
<div id="Week7">
 +
<div class="aaa"></div>
 +
<h3>Week 07 (2017/08/13~2017/08/19)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Amplify the IDT plasmid of lactate detection system</p>
 +
<style type="text/css">
 +
.tg  {border-collapse:collapse;border-spacing:0;}
 +
.tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
.tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
</style>
 +
<table class="tg">
 +
  <tr>
 +
    <th class="tg-031e">IDT plasmid # 1</th>
 +
    <th class="tg-031e">LldR gene on IDT vector</th>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-031e">IDT plasmid # 2</td>
 +
    <td class="tg-031e">GFP gene on IDT vector</td>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-031e">IDT plasmid # 3</td>
 +
    <td class="tg-031e">MazEF gene on IDT vector</td>
 +
  </tr>
 +
</table>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Amplify the IDT plasmid of CSP detection system</p>
 +
<style type="text/css">
 +
.tg  {border-collapse:collapse;border-spacing:0;}
 +
.tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
.tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
</style>
 +
<table class="tg">
 +
  <tr>
 +
    <th class="tg-031e">IDT plasmid # 1</th>
 +
    <th class="tg-031e">ComE gene on IDT vector</th>
 +
  </tr>
 +
</table>
 
</div>
 
</div>
  
−
<div id="OD600-Material">
+
<div id="Week8">
−
  <div class="aaa"></div>
+
<div class="aaa"></div>
−
<h3>Material</h3>
+
<h3>Week 08 (2017/08/20~2017/08/26)</h3>
−
<p>
+
<ol><li>Lactate detection system</li></ol>
−
1 ml LUDOX</br>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Restriction map analysis of IDT Plasmid</p>
−
mQH<sub>2</sub>O</br>
+
<ol><li>CSP detection system</li></ol>
−
96 well cell culture plate (clear with flat-bottom)
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Restriction map analysis of <i>B. subtilis</i> backbone plasmid</p>
−
</p>
+
</div>
  
−
</div>
 
  
−
<div id="OD600-Method">
+
<div id="Week9">
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
                <h3>Method</h3>
+
<h3>Week 09 (2017/08/27~2017/09/02)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Function test of kill switch (mazEF)</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Transformation test of pSBBs1C and pSBBs4S</p>
 +
</div>
  
 +
<div id="Week10">
 +
<div class="aaa"></div>
 +
<h3>Week 10 (2017/09/03~2017/09/09)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Amplify the backbone plasmid pSB1A2</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Transformation test of pSBBs1C and pSBBs4S<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Amplify the IDT plasmid of CSP detection system
 +
</p>
 +
<style type="text/css">
 +
.tg  {border-collapse:collapse;border-spacing:0;}
 +
.tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
.tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;}
 +
</style>
 +
<table class="tg">
 +
  <tr>
 +
    <th class="tg-031e">IDT plasmid # 2</th>
 +
    <th class="tg-031e">ComD gene on IDT plasmid</th>
 +
  </tr>
 +
  <tr>
 +
    <td class="tg-031e">IDT plasmid # 3</td>
 +
    <td class="tg-031e">GFP gene on IDT plasmid</td>
 +
  </tr>
 +
</table>
 +
<ol><li>Test paper experiment</li></ol>
 
<p>
 
<p>
−
1. Add 100 μl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette)</br>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Small scale culture with test paper<br/>
−
2. Add 100 μl of H<sub>2</sub>O into wells A2, B2, C2, D2 (or 1 mL H<sub>2</sub>O into cuvette)</br>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Freeze dry the test paper
−
3. Measure absorbance 600 nm of all samples in all standard measurement modes in instrument</br>
+
−
4. Record the data in excel and Import data into Excel ( OD600 reference point tab ) Sheet_1 provided</br>
+
 
</p>
 
</p>
−
 
 
</div>
 
</div>
  
−
<div id="OD600-Data-result">
+
 
−
<div class="aaa"></div>
+
<div id="Week11">
−
<h3>Data result</h3>
+
<div class="aaa"></div>
−
<br/>
+
<h3>Week 11 (2017/09/10~2017/09/16)</h3>
−
<img src="https://static.igem.org/mediawiki/2017/c/cd/Ee.jpeg" style="display:block; margin:auto;"><br/>
+
<ol><li>Lactate detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Restriction map analysis of pSB1A2</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Restriction map analysis of IDT plasmid</p>
 +
<ol><li>InterLab study</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Construct OD600 reference point<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Construct fluorescence standard curve
 +
</p>
 
</div>
 
</div>
−
</section>
 
−
 
−
<section>
 
−
 
−
 
−
<div id="Cell">
 
−
<h2>Cell measure</h2>
 
  
 +
<div id="Week12">
 +
<div class="aaa"></div>
 +
<h3>Week 12 (2017/09/17~2017/09/23)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pSB1A2_GFP_LldR assembly<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pSB1C3_mazEF assembly
 +
</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pSBBS1C_comE assembly</p>
 +
<ol><li>InterLab study</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;OD600 and fluorescence measurement of cell</p>
 
</div>
 
</div>
  
−
 
+
<div id="Week13">
−
<div id="Cell-Material">
+
−
  <div class="aaa"></div>
+
−
<h3>Material</h3>
+
−
<p>
+
−
Competent cells ( Escherichia coli strain DH5α)</br>
+
−
LB (Luria Bertani) media</br>
+
−
Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 μg /mL)</br>
+
−
50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)</br>
+
−
Incubator at 37°C</br>
+
−
1.5 ml eppendorf tubes for sample storage</br>
+
−
Ice bucket with ice</br>
+
−
Pipettes</br>
+
−
96 well plate(cell culture 96 well plate、tissue culture testplate)</br>
+
−
Devices (from InterLab Measurement Kit):</br>
+
−
1. Negative control(BBa_R0040)</br>
+
−
2. Positive control(J23151+B0032+E0040+B0010+B0012)</br>
+
−
3. Test Device 1: J23101+I13504</br>
+
−
4. Test Device 2: J23106+I13504</br>
+
−
5. Test Device 3: J23117+I13504</br>
+
−
6. Test Device 4: J23101+BCD2+E0040+B0015</br>
+
−
7. Test Device 5: J23106+BCD2+E0040+B0015</br>
+
−
8. Test Device 6: J23117+BCD2+E0040+B0015</br>
+
−
</p>
+
−
 
+
−
</div>
+
−
 
+
−
<div id="Cell-Method">
+
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
                <h3>Method</h3>
+
<h3>Week 13 (2017/09/24~2017/09/30)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Restriction map analysis of recombination DNA</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pSBBs1C_comD assembly</p>
 +
<ol><li>InterLab study</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;E-mail InterLab data to iGEM<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Fill out three online form of InterLab
 +
</p>
 +
<ol><li>Test paper experiment</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Culture the bacteria with 600 mL LB in Erlenmeyer flask which contains test paper<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Freeze dry the test paper
 +
</p>
 +
</div>
  
 +
<div id="Week14">
 +
<div class="aaa"></div>
 +
<h3>Week 14 (2017/10/01~2017/10/07)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 
<p>
 
<p>
−
1. Day 1 : Resuspended each plasmid in plate 7 and transform into Escherichia coli DH5α.</br>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Function test of lactate detector<br/>
−
&nbsp;(Transformation protocol is from iGEM)</br>
+
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Function test of kill switch
−
2. Day 2 : Pick 2 colonies from each of plate and inoculate it on 5-10 mL LB medium +Chloramphenicol.Grow the cells overnight (16-18 hours) at 37°C and 170 rpm.</br>
+
−
3. Day 3 : Set instrument to read OD600 (as OD calibration setting)and measure OD600 of the overnight cultures</br>
+
−
4. Dilute the cultures to a target OD 600 of 0.02 in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube (covered with foil to block light).</br>
+
−
5. Incubate the cultures at 37°C and 170 rpm.</br>
+
−
6. Take 1000 μL samples of the cultures at 0, 2, 4, and 6 hours of incubation and place samples on ice</br>
+
−
7. 4 replicates of 100 uL samples were taken from each culture at 0, 2, 4, and 6 hours of incubation and placed in a 96 well plate for OD and fluorescence measurements using the setup described above</br>
+
 
</p>
 
</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pSBBs1C_comE_comD assembly<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;pSBBs4S_GFP assembly
 +
</p>
 +
<ol><li>Test paper experiment</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Culture the bacteria with 600 mL LB in Erlenmeyer flask which contains test paper<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Freeze dry the liquid culture
 +
</p>
 +
</div>
  
 +
<div id="Week15">
 +
<div class="aaa"></div>
 +
<h3>Week 15 (2017/10/08~2017/10/14)</h3>
 +
<ol><li>Lactate detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Double plasmid transformation test<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Function test of lactate detector<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Function test of kill switch
 +
</p>
 +
<ol><li>CSP detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Check the size of assembly plasmid<br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Transform the recombination plasmid to <i>B. subtilis</i>
 +
</p>
 +
<ol><li>Test paper experiment</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Revive the freeze-dried bacteria</p>
 
</div>
 
</div>
  
−
<div id="Cell-Data-result">
+
 
 +
<div id="Week16">
 
<div class="aaa"></div>
 
<div class="aaa"></div>
−
<h3>Data result</h3>
+
<h3>Week 16 (2017/10/15~2017/10/21)</h3>
−
<br/>
+
<ol><li>Lactate detection system</li></ol>
−
<img src="https://static.igem.org/mediawiki/2017/f/f7/IL_cell1.jpg" style="display:block; margin:auto;"><br/><br/>
+
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Bacteria which contain two plasmid</p>
−
<img src="https://static.igem.org/mediawiki/2017/4/41/IL_cell2.jpg" style="display:block; margin:auto;"><br/>
+
<ol><li>CSP detection system</li></ol>
 +
<p>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Prepare competent cell of <i>B. subtilis</i><br/>
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Transform the recombination plasmid to <i>B. subtilis</i>
 +
</p>
 +
<ol><li>Test paper experiment</li></ol>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Freeze dry the liquid culture, and then revive it</p>
 
</div>
 
</div>
 
</section>
 
</section>
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<a href="https://www.facebook.com/ccuigemteam" target="_blank">
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</a>
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</li>
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              <li>
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              <a href="emailto:ccu.igem.2017@gmail.com">
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              <img src="https://static.igem.org/mediawiki/2016/e/e2/T--Harvard_BioDesign--images_gmail01.png" alt="Email Logo" style="width:51px;height:51px;">
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<!-- Copyright -->
 
<!-- Copyright -->
 
<div class="copyright">
 
<div class="copyright">
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<ul class="menu">
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<li>&#169; 2017 CCU Taiwan iGEM</li><li>Design: <a href="http://html5up.net">HTML5 UP</a></li>
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Latest revision as of 01:29, 2 November 2017

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Week 01 (2017/07/02~2017/07/08)

  1. Lactate detection system

          Gene design of lactate detector

  1. CSP detection system

          Gene design of CSP detector

Week 02 (2017/07/09~2017/07/15)

  1. Lactate detection system

          Gene design of lactate detector

  1. CSP detection system

          Gene design of CSP detector
          Request 2 backbone plasmids of Bacillus subtilis from iGEM

Part requesting #1 BBa_K823022 pSBBs4S
Part requesting #2 BBa_K823023 pSBBs1C

          Order chemical materials for preparing competent cell of Bacillus subtilis

Week 03 (2017/07/16~2017/07/22)

  1. Lactate detection system

          Gene design of lactate detector

  1. CSP detection system

          Gene design of CSP detector

Week 04 (2017/07/23~2017/07/29)

  1. Lactate detection system

          Send orders to IDT for DNA synthesis

  1. CSP detection system

          Gene design of CSP detector

Week 05 (2017/07/30~2017/08/05)

  1. Participate the 5th iGEM Asia-Pacific Conference at NCTU

Week 06 (2017/08/06~2017/08/12)

  1. Lactate detection system
  1. CSP detection system

          Prepare competent cell of Bacillus subtilis
          - Amplify two backbone plasmids of Bacillus subtilis

  1. InterLab Study

          Transform eight kinds of plasmids into E. coli DH5α

Positive Control BBa_I20270
Negative Control BBa_R0040
Test Device 1 BBa_J364000
Test Device 2 BBa_J364001
Test Device 3 BBa_J364002
Test Device 4 BBa_J364003
Test Device 5 BBa_J364004
Test Device 6 BBa_J364005

          Plate culture eight kinds of E. coli DH5α

Week 07 (2017/08/13~2017/08/19)

  1. Lactate detection system

          Amplify the IDT plasmid of lactate detection system

IDT plasmid # 1 LldR gene on IDT vector
IDT plasmid # 2 GFP gene on IDT vector
IDT plasmid # 3 MazEF gene on IDT vector
  1. CSP detection system

          Amplify the IDT plasmid of CSP detection system

IDT plasmid # 1 ComE gene on IDT vector

Week 08 (2017/08/20~2017/08/26)

  1. Lactate detection system

          Restriction map analysis of IDT Plasmid

  1. CSP detection system

          Restriction map analysis of B. subtilis backbone plasmid

Week 09 (2017/08/27~2017/09/02)

  1. Lactate detection system

          Function test of kill switch (mazEF)

  1. CSP detection system

          Transformation test of pSBBs1C and pSBBs4S

Week 10 (2017/09/03~2017/09/09)

  1. Lactate detection system

          Amplify the backbone plasmid pSB1A2

  1. CSP detection system

          Transformation test of pSBBs1C and pSBBs4S
          Amplify the IDT plasmid of CSP detection system

IDT plasmid # 2 ComD gene on IDT plasmid
IDT plasmid # 3 GFP gene on IDT plasmid
  1. Test paper experiment

          Small scale culture with test paper
          Freeze dry the test paper

Week 11 (2017/09/10~2017/09/16)

  1. Lactate detection system

          Restriction map analysis of pSB1A2

  1. CSP detection system

          Restriction map analysis of IDT plasmid

  1. InterLab study

          Construct OD600 reference point
          Construct fluorescence standard curve

Week 12 (2017/09/17~2017/09/23)

  1. Lactate detection system

          pSB1A2_GFP_LldR assembly
          pSB1C3_mazEF assembly

  1. CSP detection system

          pSBBS1C_comE assembly

  1. InterLab study

          OD600 and fluorescence measurement of cell

Week 13 (2017/09/24~2017/09/30)

  1. Lactate detection system

          Restriction map analysis of recombination DNA

  1. CSP detection system

          pSBBs1C_comD assembly

  1. InterLab study

          E-mail InterLab data to iGEM
          Fill out three online form of InterLab

  1. Test paper experiment

          Culture the bacteria with 600 mL LB in Erlenmeyer flask which contains test paper
          Freeze dry the test paper

Week 14 (2017/10/01~2017/10/07)

  1. Lactate detection system

          Function test of lactate detector
          Function test of kill switch

  1. CSP detection system

          pSBBs1C_comE_comD assembly
          pSBBs4S_GFP assembly

  1. Test paper experiment

          Culture the bacteria with 600 mL LB in Erlenmeyer flask which contains test paper
          Freeze dry the liquid culture

Week 15 (2017/10/08~2017/10/14)

  1. Lactate detection system

          Double plasmid transformation test
          Function test of lactate detector
          Function test of kill switch

  1. CSP detection system

          Check the size of assembly plasmid
          Transform the recombination plasmid to B. subtilis

  1. Test paper experiment

          Revive the freeze-dried bacteria

Week 16 (2017/10/15~2017/10/21)

  1. Lactate detection system

          Bacteria which contain two plasmid

  1. CSP detection system

          Prepare competent cell of B. subtilis
          Transform the recombination plasmid to B. subtilis

  1. Test paper experiment

          Freeze dry the liquid culture, and then revive it