Difference between revisions of "Team:Amoy/Notebook"
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<a style="width:50px; height:50px; position:fixed; right:30px; bottom:30px; z-index:9999; display:none; background:url(https://static.igem.org/mediawiki/2015/c/c1/Amoy-Home_Up_pic.gif) no-repeat center center #147abc;" href="javascript:;" class="lanrenzhijia_top"></a> | <a style="width:50px; height:50px; position:fixed; right:30px; bottom:30px; z-index:9999; display:none; background:url(https://static.igem.org/mediawiki/2015/c/c1/Amoy-Home_Up_pic.gif) no-repeat center center #147abc;" href="javascript:;" class="lanrenzhijia_top"></a> | ||
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<p class="detail_p">Extract plasmid from dry powder</br></p> | <p class="detail_p">Extract plasmid from dry powder</br></p> | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
| − | <p class="detail_p">1. Add | + | <p class="detail_p">1. Add 20μl ddH<sub>2</sub>O to solve the dry powder.</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; 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> | <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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm.</br> |
| − | 5. Plasmid | + | 5. Plasmid extraction.</br></p> |
| − | <p class="detail_h1">Product:</br></p> | + | <p class="detail_h1">Product: </br></p> |
| − | <p class="detail_p"> | + | <p class="detail_p"> Plasmid of promoter_J23100</br></br></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 93: | Line 96: | ||
<p class="detail_p">Extract plasmid from dry powder</br></p> | <p class="detail_p">Extract plasmid from dry powder</br></p> | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
| − | <p class="detail_p">1. Add | + | <p class="detail_p">1. Add 20μl ddH<sub>2</sub>O to solve the dry powder</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/d/d9/Amoy-Notebook_node42-1.jpeg" /> |
<p class="detail_p"></br>3. Pick a single colony from the agar plate using a sterile pipette tip. </br> | <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 ampicillin for 12h</br> | + | 4. Culture at 10ml LB of ampicillin for 12h, 37℃, 200rpm.</br> |
| − | 5. Plasmid | + | 5. Plasmid extraction.</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Plasmid of promoter_LacI</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 111: | Line 114: | ||
<p class="detail_p">Extract plasmid from dry powder</br></p> | <p class="detail_p">Extract plasmid from dry powder</br></p> | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
| − | <p class="detail_p">1. Add | + | <p class="detail_p">1. Add 20μl ddH<sub>2</sub>O to solve the dry powder</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/6/64/Amoy-Notebook_node43-1.JPG" /> |
<p class="detail_p"></br>3. Pick a single colony from the agar plate using a sterile pipette tip. </br> | <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 ampicillin for 12h</br> | + | 4. Culture at 10ml LB of ampicillin for 12h, 37℃, 200rpm</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of RBS_B0034</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 130: | Line 133: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μl ddH<sub>2</sub>O to solve the dry powder</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/cd/Amoy-Notebook_node44-1.jpeg" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </br> | 3. Pick a single colony from the agar plate using a sterile pipette tip. </br> | ||
| − | 4. Culture at 10ml LB of ampicillin for 12h</br> | + | 4. Culture at 10ml LB of ampicillin for 12h, 37℃, 200rpm</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of RBS_B0032</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 150: | Line 153: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μl ddH<sub>2</sub>O to solve the dry powder</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/e/e4/Amoy-Notebook_node45-1.jpeg" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of RBS_B0030</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 170: | Line 173: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μL ddH<sub>2</sub>O to solve the dry powder.</br> |
| − | 2. Suck | + | 2. Suck 10μL of plasmid into 50μL of competent cell for transformation.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/cf/Amoy-Notebook_node33-1.JPG" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm.</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of Plac_RBS_B0034</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 190: | Line 193: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μL ddH<sub>2</sub>O to solve the dry powder.</br> |
| − | 2. Suck | + | 2. Suck 10μL of plasmid into 50μL of competent cell for transformation.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/1a/Amoy-Notebook_node34-1.JPG" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm.</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of P<sub>J23100</sub>_RBS_B0030</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 210: | Line 213: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μl ddH<sub>2</sub>O to solve the dry powde</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/d/d5/Amoy-Notebook_node48-1.JPG" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of terminator_B0015</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 230: | Line 233: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μl ddH<sub>2</sub>O to solve the dry powde</br> |
| − | 2. Suck | + | 2. Suck 10μl of plasmid into 50μl of competent cell for transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/5/5c/Amoy-Notebook_node49-1.jpeg" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of terminator_B1006</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 247: | Line 250: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">PCR amplification for gene_<i>leudh</i> and double enzyme digestion</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. | + | 1. PCR amplification system for gene_<i>leudh</i></br></p> |
<table class="col-md-12" style="margin-bottom: 50px;"> | <table class="col-md-12" style="margin-bottom: 50px;"> | ||
| Line 259: | Line 262: | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">ddH<sub>2</sub>O</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 16μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"><i>leudh</i>-F</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 1.5μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"><i>leudh</i>-R</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 1.5μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">pUB18-<i>leudh</i></td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 1μL</td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6"> | + | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6">Prime Star</td> |
| − | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> 20μL </td> |
</tr class="col-md-3"> | </tr class="col-md-3"> | ||
</table> | </table> | ||
| − | <p class="detail_p">2. | + | <p class="detail_p">2. Electrophoresis of PCR products of gene_<i>leudh</i></br> |
| − | 3. Double enzyme digestion of | + | 3. Double enzyme digestion of gene_<i>leudh</i></br></p> |
<table class="col-md-12" style="margin-bottom: 50px;"> | <table class="col-md-12" style="margin-bottom: 50px;"> | ||
<tr style="border-bottom: 1px solid #aaaaaa; border-top: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa; border-top: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">Components</td> |
| − | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa;" class="col-md-6">Volume</td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> Gene </td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 20μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">ddH<sub>2<sub>O</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 20μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
<td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">buffer</td> | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">buffer</td> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 10μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">EcoR I</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 5μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6"> | + | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6">Spe I</td> |
| − | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> 5μL </td> |
</tr class="col-md-3"> | </tr class="col-md-3"> | ||
</table> | </table> | ||
| − | <p class="detail_p">4. Cycle purity of digested | + | <p class="detail_p">4. Cycle purity of digested gene_<i>leudh</i></br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Digested gene_<i>leudh</i> </br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 330: | Line 333: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">PCR amplification for gene_<i>fdh</i> and double enzyme digestion</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. | + | 1. PCR amplification system for gene_<i>fdh</i></br></p> |
<table class="col-md-12" style="margin-bottom: 50px;"> | <table class="col-md-12" style="margin-bottom: 50px;"> | ||
| Line 342: | Line 345: | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">ddH<sub>2</sub>O</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 16μL</td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"><i>fdh</i>-F</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 1.5μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"><i>fdh</i>-R</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 1.5μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">pUB18-<i>fdh</i></td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 1μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6"> | + | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6">Prime Star</td> |
| − | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> 20μL </td> |
</tr class="col-md-3"> | </tr class="col-md-3"> | ||
</table> | </table> | ||
| − | <p class="detail_p">2. | + | <p class="detail_p">2. Electrophoresis of PCR products of gene_<i>fdh</i></br> |
| − | 3. Double enzyme digestion of | + | 3. Double enzyme digestion of gene_<i>fdh</i></br></p> |
<table class="col-md-12" style="margin-bottom: 50px;"> | <table class="col-md-12" style="margin-bottom: 50px;"> | ||
<tr style="border-bottom: 1px solid #aaaaaa; border-top: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa; border-top: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">Components</td> |
| − | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-bottom: 10px; border-right: 1px solid #aaaaaa;" class="col-md-6">Volume</td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> Gene </td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 50μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">ddH<sub>2</sub>O</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6">10μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
<td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">buffer</td> | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">buffer</td> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 10μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6"> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa;" class="col-md-6">EcoR I</td> |
| − | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="padding-top: 15px; border-right: 1px solid #aaaaaa;" class="col-md-6"> 5μL </td> |
</tr> | </tr> | ||
<tr style="border-bottom: 1px solid #aaaaaa;"> | <tr style="border-bottom: 1px solid #aaaaaa;"> | ||
| − | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6"> | + | <td style="border-right: 1px solid #aaaaaa; border-left: 1px solid #aaaaaa; " class="col-md-6">Spe I</td> |
| − | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> </td> | + | <td style="border-right: 1px solid #aaaaaa;" class="col-md-6"> 5μL </td> |
</tr class="col-md-3"> | </tr class="col-md-3"> | ||
</table> | </table> | ||
| − | <p class="detail_p">4. Cycle purity of digested | + | <p class="detail_p">4. Cycle purity of digested gene_<i>fdh</i></br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Digested gene_<i>fdh</i></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 413: | Line 416: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of gene_<i>leudh</i> and terminator</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of gene_<i>leudh</i> and terminator_B0015</br> |
| − | 2. Electrophoresis analysis of double digested result of plasmid</br> | + | 2. Electrophoresis analysis of double digested result of plasmid.</br> |
| − | 3. Extract double digested | + | 3. Extract double digested gene_<i>leudh</i>.</br> |
| − | 4. Cycle purity of digested terminator</br> | + | 4. Cycle purity of digested terminator.</br> |
| − | 5. | + | 5. Ligate under 16℃ for 8 hours.</br> |
6. Transformation</br></p> | 6. Transformation</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/3/34/Amoy-Notebook_node35-1.jpeg" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol</br> | + | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 8. Extract the plasmids</br> | + | 8. Extract the plasmids.</br> |
| − | 9. Electrophoresis analysis of plasmids</br></p> | + | 9. Electrophoresis analysis of plasmids.</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Plasmid of gene_<i>leudh</i> and terminator_B0015</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 437: | Line 440: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of gene_<i>fdh</i> and terminator</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of gene_<i>fdh</i> and terminator_B0015</br> |
| − | 2. Electrophoresis analysis of double digested result of plasmid</br> | + | 2. Electrophoresis analysis of double digested result of plasmid.</br> |
| − | 3. Extract double digested | + | 3. Extract double digested gene_<i>fdh</i>.</br> |
| − | 4. Cycle purity of digested terminator</br> | + | 4. Cycle purity of digested terminator.</br> |
| − | 5. | + | 5. Ligate under 16℃ for 8 hours.</br> |
6. Transformation</br></p> | 6. Transformation</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/0/01/Amoy-Notebook_Node36_figure1.JPG" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol</br> | + | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 8. Extract the plasmids</br> | + | 8. Extract the plasmids.</br> |
| − | 9. Electrophoresis analysis of plasmids</br></p> | + | 9. Electrophoresis analysis of plasmids.</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Plasmid of gene_<i>fdh</i> and terminator_B0015</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 461: | Line 464: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of gene_<i>leudh</i> and terminator</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of gene_<i>leudh</i> and terminator_B1006</br> |
| − | 2. Electrophoresis analysis of double digested result of plasmid</br> | + | 2. Electrophoresis analysis of double digested result of plasmid.</br> |
| − | 3. Extract double digested | + | 3. Extract double digested gene_<i>leudh</i> and terminator.</br> |
| − | + | 4. Ligate under 16℃ for 8 hours.</br> | |
| − | + | 5. Transformation</br></p> | |
| − | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c1/Amoy-Notebook_node37-1.jpeg" /> | |
| − | <img style="width: | + | |
<p class="detail_p"></br> | <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. Culture at 37℃,200rpm for 12~14 hours.</br> | |
| − | + | 7. Extract the plasmids.</br> | |
| − | + | 8. Electrophoresis analysis of plasmids.</br></p> | |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Plasmid of gene_<i>leudh</i> and terminator_B1006</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 485: | Line 487: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of gene_<i>fdh</i> and terminator</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of gene_<i>fdh</i> and terminator_B1006</br> |
| − | 2. Electrophoresis analysis of double digested result of plasmid</br> | + | 2. Electrophoresis analysis of double digested result of plasmid.</br> |
| − | 3. Extract double digested | + | 3. Extract double digested gene_<i>fdh</i> and terminator.</br> |
| − | + | 4. Ligate under 16℃ for 8 hours.</br> | |
| − | + | 5. Transformation</br></p> | |
| − | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/18/Amoy-Notebook_node38-1.jpeg" /> | |
| − | <img style="width: | + | |
<p class="detail_p"></br> | <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. Culture at 37℃,200rpm for 12~14 hours.</br> | |
| − | + | 7. Extract the plasmids.</br> | |
| − | + | 8. Electrophoresis analysis of plasmids.</br></p> | |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Plasmid of gene_<i>fdh</i> and terminator_B1006</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 509: | Line 510: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of isolated circuit with RBS_B0032 and gene_<i>leudh</i></br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of Plac_B0032 and LeuDH_T.</br> |
| − | 2. Electrophoresis analysis of double digested result | + | 2. Electrophoresis analysis of double digested result.</br> |
| − | + | 3. Extract double digested products.</br> | |
| − | + | 4. Ligate under 16℃ for 8 hours.</br> | |
| − | 3. Extract double digested | + | 5. Transformation</br> |
| − | 4. | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/5/57/Amoy-Notebook_node21-2.JPG" /> |
| − | 5 | + | |
| − | + | ||
| − | <img style="width: | + | |
<p class="detail_p"></br> | <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. Culture at 37℃,200rpm for 12~14 hours.</br> | |
| − | + | 7. Extract the plasmids.</br> | |
| − | + | 8. Electrophoresis analysis of plasmids.</br></p> | |
| − | + | ||
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Isolated circuit with RBS_B0032 and gene_<i>leudh</i></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 536: | Line 533: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of isolated circuit with RBS_B0034 and gene_<i>leudh</i></br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of Plac_B0034 and LeuDH_TT</br> |
| − | 2. Electrophoresis analysis of double digested result</br></p> | + | 2. Electrophoresis analysis of double digested result.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/e/e2/Amoy-Notebook_Node2_figure1.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 3. Extract double digested | + | 3. Extract double digested products.</br> |
| − | 4. | + | 4. Ligate under 16℃ for 8 hours.</br> |
| − | 5 | + | 5. Transformation</br></p> |
| − | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/4/43/Amoy-Notebook_node24-2.JPG" /> | |
| − | <img style="width: | + | |
<p class="detail_p"></br> | <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. Culture at 37℃,200rpm for 12~14 hours.</br> | |
| − | + | 7. Extract the plasmids</br> | |
| − | + | 8. Electrophoresis analysis of plasmids</br></p> | |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/e/e3/Amoy-Notebook_Node22_figure3.png" /> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Isolated circuit with RBS_B0034 and gene_<i>leudh</i></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 563: | Line 559: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of isolated circuit with RBS_B0034 and gene_<i>fdh</i></br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of Plac_B0032 and LeuDH_T</br> |
2. Electrophoresis analysis of double digested result</br></p> | 2. Electrophoresis analysis of double digested result</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/e/e2/Amoy-Notebook_Node2_figure1.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 3. Extract double digested | + | 3. Extract double digested products.</br> |
| − | 4. | + | 4. Ligate under 16℃ for 8 hours.</br> |
| − | 5 | + | 5. Transformation</br></p> |
| − | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/3/3e/Amoy-Notebook_node23-2.JPG" /> | |
| − | <img style="width: | + | |
<p class="detail_p"></br> | <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. Culture at 37℃,200rpm for 12~14 hours.</br> | |
| − | + | 7. Extract the plasmids.</br> | |
| − | + | 8. Electrophoresis analysis of plasmids.</br></p> | |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/d/d7/Amoy-Notebook_Node23_figure3.png" /> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Isolated circuit with RBS_B0034 and gene_<i>fdh</i></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 590: | Line 585: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of isolated circuit with RBS_B0030 and gene_<i>leudh</i></br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of P<sub>J23100</sub>_B0030 and LeuDH_T</br> |
| − | 2. Electrophoresis analysis of double digested result | + | 2. Electrophoresis analysis of double digested result.</br> |
| − | + | 3. Extract double digested products.</br> | |
| − | + | 4. Ligate under 16℃ for 8 hours.</br> | |
| − | 3. Extract double digested | + | 5. Transformation</br> |
| − | 4. | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/f/f8/Amoy-Notebook_Node24_figure2.JPG" /> |
| − | 5 | + | |
| − | + | ||
| − | <img style="width: | + | |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol</br> | + | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 8. Extract the plasmids</br> | + | 8. Extract the plasmids.</br> |
| − | 9. Electrophoresis analysis of plasmids</br></p> | + | 9. Electrophoresis analysis of plasmids.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c9/Amoy-Notebook_Node24_figure3.png" /> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Isolated circuit with RBS_B0030 and gene_<i>leudh</i></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 617: | Line 609: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of promoter and RBS_B0032</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of P<sub>lac</sub> and RBS_B0032</br> |
2. Electrophoresis analysis of double digested result of plasmid</br></p> | 2. Electrophoresis analysis of double digested result of plasmid</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/5/50/Amoy-Notebook_Node42_figure1.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 3. Extract double digested | + | 3. Extract double digested Plac</br> |
| − | 4. Cycle purity of digested | + | 4. Cycle purity of digested RBS_B0032</br> |
| − | 5. | + | 5. Ligate under 16℃ for 8 hours.</br> |
| − | 6. | + | 6. Transformation</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/7/7a/Amoy-Notebook_node32-2.jpeg" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 7. Pick | + | 7. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 8. Extract the plasmids</br> | + | 8. Extract the plasmids.</br> |
| − | 9. Electrophoresis analysis of plasmids</br></p> | + | 9. Electrophoresis analysis of plasmids.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/f/fa/Amoy-Notebook_Node42_figure3.png" /> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of Plac linked with RBS_B0032</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 644: | Line 636: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of the final circuit with RBS_B0032</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of circuits with RBS_B0032_gene_<i>leudh</i> and RBS_B0034_gene_<i>fdh</i>. </br> |
| − | 2. Electrophoresis analysis of double digested result</br></p> | + | 2. Electrophoresis analysis of double digested result.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/2/2c/Amoy-Notebook_Node11_figure1.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 3. Extract double digested | + | 3. Extract double digested products.</br> |
| − | 4. | + | 4. Ligate under 16℃ for 8 hours.</br> |
5. Transformation</br></p> | 5. Transformation</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/b/b9/Amoy-Notebook_node11-2.JPG" /> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 9. Verify the results by double enzyme digestion</br></p> | + | 6. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | <img style="width: | + | 7. Extract the plasmids.</br> |
| + | 8. Electrophoresis analysis of plasmids.</br> | ||
| + | 9. Verify the results by double enzyme digestion.</br></p> | ||
| + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/8/80/Amoy-Notebook_Node11_figure3.png" /> | ||
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Final circuit of RBS_B0032</br></br></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 673: | Line 663: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of the final circuit with RBS_B0034</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of circuit with | + | 1. Double enzyme digestion of circuit with RBS_B0034_gene_<i>leudh</i> and RBS_B0034_gene_<i>fdh</i>.</br> |
| − | 2. Electrophoresis analysis of double digested result</br></p> | + | 2. Electrophoresis analysis of double digested result.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right:40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/5/51/Amoy-Notebook_Node12_figure1.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 3. Extract double digested | + | 3. Extract double digested products.</br> |
| − | 4. | + | 4. Ligate under 16℃ for 8 hours.</br> |
5. Transformation</br></p> | 5. Transformation</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/0/01/Amoy-Notebook_node12-2.jpeg" /> |
<p class="detail_p"></br> | <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> | + | 6. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 7. Extract the plasmids</br> | + | 7. Extract the plasmids.</br> |
| − | 8. Electrophoresis analysis of plasmids</br></p> | + | 8. Electrophoresis analysis of plasmids.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/4/4e/Amoy-Notebook_Node12_figure3.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 9. Verify the results by double enzyme digestion</br></p> | + | 9. Verify the results by double enzyme digestion.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/9/9f/Amoy-Notebook_Node12_figure4.png" /> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Final circuit of RBS_B0034</br></br></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 702: | Line 692: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of the final circuit with RBS_B0030</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of circuit with | + | 1. Double enzyme digestion of circuit with RBS_B0030_gene_<i>leudh</i> and RBS_B0034_gene_<i>fdh</i>.</br> |
2. Electrophoresis analysis of double digested result</br></p> | 2. Electrophoresis analysis of double digested result</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/a/ae/Amoy-Notebook_Node13_figure1.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 3. Extract double digested | + | 3. Extract double digested products.</br> |
| − | 4. | + | 4. Ligate under 16℃ for 8 hours.</br> |
5. Transformation</br></p> | 5. Transformation</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/18/Amoy-Notebook_node13-2.JPG" /> |
<p class="detail_p"></br> | <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> | + | 6. Pick 8 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 7. Extract the plasmids</br> | + | 7. Extract the plasmids.</br> |
| − | 8. Electrophoresis analysis of plasmids</br></p> | + | 8. Electrophoresis analysis of plasmids.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/2/29/Amoy-Notebook_Node13_figure3.png" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 9. Verify the results by double enzyme digestion</br></p> | + | 9. Verify the results by double enzyme digestion.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/c7/Amoy-Notebook_Node13_figure4.png" /> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Final circuit of RBS_B0030</br></br></br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 734: | Line 724: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μL ddH<sub>2</sub>O to solve the dry powder.</br> |
| − | 2. Suck | + | 2. Suck 10μL of plasmid into 50μL of competent cell for transformation.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/c/cf/Amoy-Notebook_node33-1.JPG" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm.</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of Plac_RBS_B0034</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 754: | Line 744: | ||
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Add | + | 1. Add 20μL ddH<sub>2</sub>O to solve the dry powder.</br> |
| − | 2. Suck | + | 2. Suck 10μL of plasmid into 50μL of competent cell for transformation.</br></p> |
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/1a/Amoy-Notebook_node34-1.JPG" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
3. Pick a single colony from the agar plate using a sterile pipette tip. </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> | + | 4. Culture at 10ml LB of chloramphenicol for 12h, 37℃, 200rpm.</br> |
| − | 5. Plasmid | + | 5. Plasmid Extraction</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p"> </br></p> | + | <p class="detail_p"> Plasmid of P<sub>J23100</sub>_RBS_B0030</br></p> |
</div> | </div> | ||
</div> | </div> | ||
| Line 771: | Line 761: | ||
<div style="width: 80%; margin-left: 10%;"> | <div style="width: 80%; margin-left: 10%;"> | ||
<p class="detail_h1">Purpose:</br></p> | <p class="detail_h1">Purpose:</br></p> | ||
| − | <p class="detail_p"> | + | <p class="detail_p">Ligation of promoter and RBS</br></p> |
<p class="detail_h1">Steps:</br></p> | <p class="detail_h1">Steps:</br></p> | ||
<p class="detail_p"> | <p class="detail_p"> | ||
| − | 1. Double enzyme digestion of | + | 1. Double enzyme digestion of P<sub>J23100</sub> and RBS_B0034.</br> |
| − | 2. Cycle purity of digested products</br> | + | 2. Cycle purity of digested products.</br> |
| − | 3. | + | 3. Ligate under 16℃ for 8 hours.</br> |
4. Transformation</br></p> | 4. Transformation</br></p> | ||
| − | <img style="width: | + | <img style="width: 60%; margin-right: 40%; margin-top: 10px; margin-bottom: 0px;" src="https://static.igem.org/mediawiki/2015/1/18/Amoy-Notebook_node31-1.jpeg" /> |
<p class="detail_p"></br> | <p class="detail_p"></br> | ||
| − | 5. Pick 10 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of | + | 5. Pick 10 single colonies from the agar plate using sterile pipette tips. Put it into 10ml LB of chloramphenicol. Culture at 37℃,200rpm for 12~14 hours.</br> |
| − | 6. Extract the plasmids</br> | + | 6. Extract the plasmids.</br> |
| − | 7. Electrophoresis analysis of plasmids</br></p> | + | 7. Electrophoresis analysis of plasmids.</br></p> |
<p class="detail_h1">Product:</br></p> | <p class="detail_h1">Product:</br></p> | ||
| − | <p class="detail_p">Plasmid of | + | <p class="detail_p">Plasmid of P<sub>J23100</sub> linked with RBS_B0034</br></p> |
</div> | </div> | ||
</div> | </div> | ||
<span id="node31_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="node31_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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| − | < | + | <span class="preload1"></span> <span class="preload2"></span> |
| − | + | <ul id="nav"> | |
| − | + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Team" id="products" class="top_link"><span class="down">TEAM</span></a> | |
| − | + | <ul class="sub"> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Team#member_student">Member</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Member/Amoy">Amoy</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Attributions">Attributions</a></li> | |
| − | + | </ul> | |
| − | + | </li> | |
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| − | + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Project" id="products" class="top_link"><span class="down">PROJECT</span></a> | |
| − | <ul> | + | <ul class="sub"> |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Project/Background">Background</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Description">Description</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Project/Methods">Methods</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Project/Results">Results</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Project/Discussion">Discussion</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Project/FutureWork">Future Work</a></li> | |
| − | + | </ul> | |
| − | + | </li> | |
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| − | + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Notebook" id="products" class="top_link"><span class="down active">NOTEBOOK</span></a> | |
| − | + | <ul class="sub"> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Notebook">Notebook</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Protocol">Protocols</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Parts">Parts</a></li> | |
| − | + | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Gallery">Gallery</a></li> | |
| − | + | </ul> | |
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| − | + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Interlab" id="products" class="top_link"><span class="down">INTERLAB</span></a></li> | |
| − | + | ||
| + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Newsletter" id="products" class="top_link"><span class="down">NEWSLETTER</span></a> | ||
| + | <ul class="sub"> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Newsletter#title">Introduction</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Newsletter/Contribution">Contribution</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Newsletter/Discussion">Discussion</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Newsletter/Links">Links</a></li> | ||
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| + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Practices" id="products" class="top_link"><span class="down">PRACTICES</span></a> | ||
| + | <ul class="sub"> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Practices/Promotion">Promotion</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Practices/Talk">Talk</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Practice/Communication">Communication</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Collaborations">Collaborations</a></li> | ||
| + | </ul> | ||
| + | </li> | ||
| + | |||
| + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Judging" id="products" class="top_link"><span class="down">JUDGING</span></a> | ||
| + | <ul class="sub"> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Judging/Medal">Medal Criteria</a></li> | ||
| + | <li><a href="https://2015.igem.org/Team:Amoy/Judging/Acknowledgement">Acknowledgement</a></li> | ||
| + | </ul> | ||
| + | </li> | ||
| + | |||
| + | <li class="top"><a href="https://2015.igem.org/Team:Amoy/Safety" id="products" class="top_link"><span class="down">SAFETY</span></a></li> | ||
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<ul class="ul_menu"> | <ul class="ul_menu"> | ||
<li><a href="https://2015.igem.org/Team:Amoy/Notebook/Notebook">Notebook</a></li> | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Notebook">Notebook</a></li> | ||
| − | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Protocol"> | + | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Protocol">Protocols</a></li> |
<li><a href="https://2015.igem.org/Team:Amoy/Parts">Parts</a></li> | <li><a href="https://2015.igem.org/Team:Amoy/Parts">Parts</a></li> | ||
<li><a href="https://2015.igem.org/Team:Amoy/Notebook/Gallery">Gallery</a></li> | <li><a href="https://2015.igem.org/Team:Amoy/Notebook/Gallery">Gallery</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
| − | |||
| − | |||
| − | |||
<!--word--> | <!--word--> | ||
| − | <div id="title" | + | <div id="title" style="width: 70%; margin-left: 25%;"> |
<p id="title_p">NOTEBOOK</p> | <p id="title_p">NOTEBOOK</p> | ||
| − | <p class="main_p">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 | + | <p class="main_p">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-<i>tert</i>-leucine production. So scientists introduced whole-cell biocatalysts to L-<i>tert</i>-leucine production. Whole-cell biocatalysts could stabilize enzymes and reduce the addition level of cofactor NADH.</br></br> |
| − | In the path of building our biobricks, we divided the circuits into two modules. One is promoter linked with | + | 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.</br></br> |
</p> | </p> | ||
| − | <div style="width: 800px; height: 400px;"> | + | <div style="width: 800px; height: 400px; margin-left: -20px;"> |
<svg width="800" height="400" style="position: absolute; z-index: -10;"> | <svg width="800" height="400" style="position: absolute; z-index: -10;"> | ||
<g transform="translate(0,0)"> | <g transform="translate(0,0)"> | ||
| Line 923: | Line 914: | ||
</svg> | </svg> | ||
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| − | <a id="node12" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 385px; margin-top: 35px; background-color: green; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node12" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 385px; margin-top: 35px; background-color: green; border: 0px;" data-trigger="hover" data-content="Plac_B0034_LeuDH_TT_LacI_B0034_FDH_TT" ></a> |
| − | <a id="node13" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 485px; margin-top: 35px; background-color: green; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node13" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 485px; margin-top: 35px; background-color: green; border: 0px;" data-trigger="hover" data-content="P<sub>J23100</sub>_B0030_LeuDH_T_LacI_B0034_FDH_TT" ></a> |
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| − | <a id="node23" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 435px; margin-top: 135px; background-color: yellow; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node23" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 435px; margin-top: 135px; background-color: yellow; border: 0px;" data-trigger="hover" data-content="Plac_B0034_FDH_TT" ></a> |
| − | <a id="node24" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 535px; margin-top: 135px; background-color: yellow; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node24" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 535px; margin-top: 135px; background-color: yellow; border: 0px;" data-trigger="hover" data-content="P<sub>J23100</sub>_B0030_LeuDH_T" ></a> |
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| − | <a id="node32" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 135px; margin-top: 235px; background-color: blue; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node32" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 135px; margin-top: 235px; background-color: blue; border: 0px;" data-trigger="hover" data-content="Plac_B0032" ></a> |
| − | <a id="node33" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 185px; margin-top: 235px; background-color: blue; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node33" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 185px; margin-top: 235px; background-color: blue; border: 0px;" data-trigger="hover" data-content="Plac_B0034" ></a> |
| − | <a id="node34" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 235px; margin-top: 235px; background-color: blue; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node34" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 235px; margin-top: 235px; background-color: blue; border: 0px;" data-trigger="hover" data-content="P<sub>J23100</sub>_B0030" ></a> |
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| − | <a id="node41" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 35px; margin-top: 335px; background-color: pink; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node41" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 35px; margin-top: 335px; background-color: pink; border: 0px;" data-trigger="hover" data-content="Promoter</br>BBa_J23100" ></a> |
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| − | <a id="node45" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 285px; margin-top: 335px; background-color: DarkBlue; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node45" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 285px; margin-top: 335px; background-color: DarkBlue; border: 0px;" data-trigger="hover" data-content="RBS</br>BBa_B0030" ></a> |
| − | <a id="node46" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 385px; margin-top: 335px; background-color: blue; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node46" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 385px; margin-top: 335px; background-color: blue; border: 0px;" data-trigger="hover" data-content="Plac_B0034" ></a> |
| − | <a id="node47" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 435px; margin-top: 335px; background-color: blue; border: 0px;" data-trigger="hover" data-content=" | + | <a id="node47" class="btn btn-success" rel="popover" style="border-radius: 100px; width: 30px; height: 30px; position: absolute; margin-left: 435px; margin-top: 335px; background-color: blue; border: 0px;" data-trigger="hover" data-content="P<sub>J23100</sub>_B0030" ></a> |
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Latest revision as of 03:14, 19 September 2015
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
