Difference between revisions of "Team:Amoy/Notebook"

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<span id="node45_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
 
<span id="node45_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
  
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<div id="node46_detail" style="width:80%; margin-left: 10%; height: 600px; margin-top: 40px; background-color: #fff; border-radius: 10px; border: 1px solid #aaaaaa; position: fixed; z-index: 1001; overflow: auto; display: none; padding-top: 50px; padding-bottom: 50px;">
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<div style="width: 80%; margin-left: 10%;">
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<p class="detail_h1">Purpose:</br></p>
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<p class="detail_p">Extract plasmid from dry powder</br></p>
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<p class="detail_h1">Steps:</br></p>
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<p class="detail_p">
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1. Add 20ul ddH20 to solve the dry powder</br>
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2. Suck 10ul of plasmid into 50ul of competent cell for transformation</br></p>
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<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/8/87/Amoy-Notebook_Node41_figure1.jpg" />
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<p class="detail_p"></br>
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3. Pick a single colony from the agar plate using a sterile pipette tip. </br>
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4. Culture at 10ml LB of chloramphenicol for 12h</br>
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5. Plasmid minipre</br></p>
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<p class="detail_h1">Product:</br></p>
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<p class="detail_p"> </br></p>
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</div>
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</div>
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<span id="node46_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
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<!--node47-->
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<div id="node47_detail" style="width:80%; margin-left: 10%; height: 600px; margin-top: 40px; background-color: #fff; border-radius: 10px; border: 1px solid #aaaaaa; position: fixed; z-index: 1001; overflow: auto; display: none; padding-top: 50px; padding-bottom: 50px;">
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<div style="width: 80%; margin-left: 10%;">
 +
<p class="detail_h1">Purpose:</br></p>
 +
<p class="detail_p">Extract plasmid from dry powder</br></p>
 +
<p class="detail_h1">Steps:</br></p>
 +
<p class="detail_p">
 +
1. Add 20ul ddH20 to solve the dry powde</br>
 +
2. Suck 10ul of plasmid into 50ul of competent cell for transformation</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/8/87/Amoy-Notebook_Node41_figure1.jpg" />
 +
<p class="detail_p"></br>
 +
3. Pick a single colony from the agar plate using a sterile pipette tip. </br>
 +
4. Culture at 10ml LB of chloramphenicol for 12h</br>
 +
5. Plasmid minipre</br></p>
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p"> </br></p>
 +
</div>
 +
</div>
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<span id="node47_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
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<!--node48-->
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<div id="node48_detail" style="width:80%; margin-left: 10%; height: 600px; margin-top: 40px; background-color: #fff; border-radius: 10px; border: 1px solid #aaaaaa; position: fixed; z-index: 1001; overflow: auto; display: none; padding-top: 50px; padding-bottom: 50px;">
 +
<div style="width: 80%; margin-left: 10%;">
 +
<p class="detail_h1">Purpose:</br></p>
 +
<p class="detail_p">Extract plasmid from dry powder</br></p>
 +
<p class="detail_h1">Steps:</br></p>
 +
<p class="detail_p">
 +
1. Add 20ul ddH20 to solve the dry powde</br>
 +
2. Suck 10ul of plasmid into 50ul of competent cell for transformation</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/8/87/Amoy-Notebook_Node41_figure1.jpg" />
 +
<p class="detail_p"></br>
 +
3. Pick a single colony from the agar plate using a sterile pipette tip. </br>
 +
4. Culture at 10ml LB of chloramphenicol for 12h</br>
 +
5. Plasmid minipre</br></p>
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p"> </br></p>
 +
</div>
 +
</div>
 +
<span id="node48_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
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<div id="node49_detail" style="width:80%; margin-left: 10%; height: 600px; margin-top: 40px; background-color: #fff; border-radius: 10px; border: 1px solid #aaaaaa; position: fixed; z-index: 1001; overflow: auto; display: none; padding-top: 50px; padding-bottom: 50px;">
 +
<div style="width: 80%; margin-left: 10%;">
 +
<p class="detail_h1">Purpose:</br></p>
 +
<p class="detail_p">Extract plasmid from dry powder</br></p>
 +
<p class="detail_h1">Steps:</br></p>
 +
<p class="detail_p">
 +
1. Add 20ul ddH20 to solve the dry powde</br>
 +
2. Suck 10ul of plasmid into 50ul of competent cell for transformation</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/8/87/Amoy-Notebook_Node41_figure1.jpg" />
 +
<p class="detail_p"></br>
 +
3. Pick a single colony from the agar plate using a sterile pipette tip.  </br>
 +
4. Culture at 10ml LB of chloramphenicol for 12h</br>
 +
5. Plasmid minipre</br></p>
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p"> </br></p>
 +
</div>
 +
</div>
 +
<span id="node49_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
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<div id="node50_detail" style="width:80%; margin-left: 10%; height: 600px; margin-top: 40px; background-color: #fff; border-radius: 10px; border: 1px solid #aaaaaa; position: fixed; z-index: 1001; overflow: auto; display: none; padding-top: 50px; padding-bottom: 50px;">
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<div style="width: 80%; margin-left: 10%;">
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<p class="detail_h1">Purpose:</br></p>
 +
<p class="detail_p">pcr amplification for gene ldh and double enzyme digestion</br></p>
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<p class="detail_h1">Steps:</br></p>
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<p class="detail_p">
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1. pcr amplification system for gene ldh</br></p>
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<table class="col-md-12" style="margin-bottom: 50px;">
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<tr style="border-bottom: 1px solid #aaaaaa; border-top: 1px solid #aaaaaa;">
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  <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">Components</td>
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  <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa;" class="col-md-6">Volume</td>
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</tr>
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<tr style="border-bottom: 1px solid #aaaaaa;">
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">ddH20</td>
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td>
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</tr>
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<tr style="border-bottom: 1px solid #aaaaaa;">
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">LDH-F</td>
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td>
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</tr>
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<tr style="border-bottom: 1px solid #aaaaaa;">
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">LDH-R</td>
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td>
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</tr>
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<tr style="border-bottom: 1px solid #aaaaaa;">
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">Pub18-LDH</td>
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  <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td>
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</tr>
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<tr style="border-bottom: 1px solid #aaaaaa;">
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  <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6">primestar</td>
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  <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> </td>
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</tr class="col-md-3">
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</table>
 +
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<p class="detail_p">2. electrophoresis of PCR products of gene ldh</br></p>
 +
 +
<p class="detail_p"></br>
 +
3. Double enzyme digestion of gene ldh</br>
 +
4. Cycle purity of digested gene ldh</br></p>
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p"> </br></p>
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</div>
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</div>
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<span id="node50_close" style="background: #f0f0f0 url(https://static.igem.org/mediawiki/2015/7/73/GalleryClose.gif) no-repeat center center; position: fixed; height: 18px; width: 18px; cursor: pointer; margin-top: 60px; right: 12%; z-index: 1002; display: none;"></span>
  
  

Revision as of 07:42, 11 September 2015

Aomy/Project

NOTEBOOK

Initially, they used isolated enzymes, which can be disadvantageous for the reason that enzymes are always destabilized in the isolation and purification process. What's more, the cofactor-NADH is rather an expensive raw material, which will enhance the cost of L-tert-leucine production. So scientists introduced whole-cell biocatalysts to L-tert-leucine production. Whole-cell biocatalysts could stabilize enzymes and reduce the addition level of cofactor NADH.

In the path of building our biobricks, we divided the circuits into two modules. One is promoter linked with rbs and the other is gene linked with terminator. The dendrogram below is our experiments detail. Click each bottom for more information.

CONTACT US

Email: igemxmu@gmail.com

Website: 2015.igem.org/Team:Amoy

Address: Xiamen University, No. 422, Siming South Road, Xiamen, Fujian, P.R.China 361005