Difference between revisions of "Team:Amoy/Notebook"

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</div>
 
</div>
 
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<span id="node32_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="node11_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">make connection of the final circuit with RBS_B0032</br></p>
 +
<p class="detail_h1">Steps:</br></p>
 +
<p class="detail_p">
 +
1. Double enzyme digestion of circuit with RBS_B0032 and gene_ldh and gene_fdh circuit.</br>
 +
2. Electrophoresis analysis of double digested result</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/1f/Amoy-Notebook_Node32_figure1.jpg" />
 +
<p class="detail_p"></br>
 +
3. Extract double digested product</br>
 +
4. Ligation under 16℃ for 8 hours</br>
 +
5. 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>
 +
6. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol</br>
 +
7. Extract the plasmids</br>
 +
8. Electrophoresis analysis of plasmids</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node32_figure3.jpg" />
 +
<p class="detail_p"></br>
 +
9. Verify the results by double enzyme digestion</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node32_figure3.jpg" />
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p">final circuit of RBS_B0032</br></p>
 +
</div>
 +
</div>
 +
<span id="node11_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 style="width: 80%; margin-left: 10%;">
 +
<p class="detail_h1">Purpose:</br></p>
 +
<p class="detail_p">make connection of the final circuit with RBS_B0032</br></p>
 +
<p class="detail_h1">Steps:</br></p>
 +
<p class="detail_p">
 +
1. Double enzyme digestion of circuit with RBS_B0032 and gene_ldh and gene_fdh circuit.</br>
 +
2. Electrophoresis analysis of double digested result</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/1f/Amoy-Notebook_Node32_figure1.jpg" />
 +
<p class="detail_p"></br>
 +
3. Extract double digested product</br>
 +
4. Ligation under 16℃ for 8 hours</br>
 +
5. 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>
 +
6. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol</br>
 +
7. Extract the plasmids</br>
 +
8. Electrophoresis analysis of plasmids</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node32_figure3.jpg" />
 +
<p class="detail_p"></br>
 +
9. Verify the results by double enzyme digestion</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node32_figure3.jpg" />
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p">final circuit of RBS_B0032</br></p>
 +
</div>
 +
</div>
 +
<span id="node12_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 style="width: 80%; margin-left: 10%;">
 +
<p class="detail_h1">Purpose:</br></p>
 +
<p class="detail_p">make connection of the final circuit with RBS_B0032</br></p>
 +
<p class="detail_h1">Steps:</br></p>
 +
<p class="detail_p">
 +
1. Double enzyme digestion of circuit with RBS_B0032 and gene_ldh and gene_fdh circuit.</br>
 +
2. Electrophoresis analysis of double digested result</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/1f/Amoy-Notebook_Node32_figure1.jpg" />
 +
<p class="detail_p"></br>
 +
3. Extract double digested product</br>
 +
4. Ligation under 16℃ for 8 hours</br>
 +
5. 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>
 +
6. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol</br>
 +
7. Extract the plasmids</br>
 +
8. Electrophoresis analysis of plasmids</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node32_figure3.jpg" />
 +
<p class="detail_p"></br>
 +
9. Verify the results by double enzyme digestion</br></p>
 +
<img style="width: 30%; margin-right: 70%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node32_figure3.jpg" />
 +
<p class="detail_h1">Product:</br></p>
 +
<p class="detail_p">final circuit of RBS_B0032</br></p>
 +
</div>
 +
</div>
 +
<span id="node13_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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Revision as of 06:34, 12 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