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Team:Sumbawagen/Notebook/protocol2
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<a class="brand" style="padding:0px 15px;width:70px;height:50px;" href="https://igem.org"><img width="70px" height="50px" src="https://static.igem.org/mediawiki/2013/1/17/IGEM_basic_Logo_white_stylized.png"> </a> | <a class="brand" style="padding:0px 15px;width:70px;height:50px;" href="https://igem.org"><img width="70px" height="50px" src="https://static.igem.org/mediawiki/2013/1/17/IGEM_basic_Logo_white_stylized.png"> </a> | ||
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<a href="#" class="dropdown-toggle" data-toggle="dropdown">Overviews <b class="caret"></b></a> | <a href="#" class="dropdown-toggle" data-toggle="dropdown">Overviews <b class="caret"></b></a> | ||
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| - | <li><a href="https://2014.igem.org/Team:Sumbawagen/overviews/ | + | <li><a href="https://2014.igem.org/Team:Sumbawagen/overviews/Econey_Project"> Econey Project </a></li> |
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<li><a href="https://2014.igem.org/Team:Sumbawagen/parts">Parts</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/parts">Parts</a></li> | ||
<li><a href="https://2014.igem.org/Team:Sumbawagen/Safety">Safety</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Safety">Safety</a></li> | ||
| - | <li><a href="https://2014.igem.org/Team:Sumbawagen/Attributions">Attribution</a></li> | + | <li><a href="https://2014.igem.org/Team:Sumbawagen/Attributions">Attribution</a></li> |
| + | <li><a href="https://2014.igem.org/Team:Sumbawagen/future_direction">Future Direction</a></li> | ||
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<a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook <b class="caret"></b></a> | <a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook <b class="caret"></b></a> | ||
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| - | <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook"> | + | |
| + | <li class="nav-header">Daily Notes</li> | ||
| + | <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook">Policy & Practices</a></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook2">Lab</a></li> | ||
<li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook/Protocol">Protocol</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Notebook/Protocol">Protocol</a></li> | ||
<!--<li><a href="#">Improvements</a></li>--> | <!--<li><a href="#">Improvements</a></li>--> | ||
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<li><a href="https://2014.igem.org/Team:Sumbawagen/Team">Meet the Team</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Team">Meet the Team</a></li> | ||
| - | |||
<li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Gallery">Gallery</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Gallery">Gallery</a></li> | ||
<li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Contact">Contact</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Team/Contact">Contact</a></li> | ||
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<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/moyo">Moyo Festival</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/moyo">Moyo Festival</a></li> | ||
<li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/virtual">Virtual Outreach</a></li> | <li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/virtual">Virtual Outreach</a></li> | ||
| - | <li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/Support"> | + | <li><a href="https://2014.igem.org/Team:Sumbawagen/Human_practice/Support">Responses and supports</a></li> |
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<div class="span12"> | <div class="span12"> | ||
| - | <h2> | + | <h2>Notebook – Protocol – 2.PCR (Polymerase Chain Reaction)</h2> |
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<li>10 uM reverse primer, 5 ul</li> | <li>10 uM reverse primer, 5 ul</li> | ||
<li>Sterilized distilled water (Akuabides), 15 ul</li> | <li>Sterilized distilled water (Akuabides), 15 ul</li> | ||
| - | <li>10 uM forward primer, 5 ul</li | + | <li>10 uM forward primer, 5 ul</li> |
<li>DNA template, colony from agar plate 0 ul</li> | <li>DNA template, colony from agar plate 0 ul</li> | ||
| - | Total volume, 50 ul | + | Total volume, 50 ul</p></ul> |
<p>4.Reaction conditions for PCR were as follow:<br> | <p>4.Reaction conditions for PCR were as follow:<br> | ||
Latest revision as of 09:11, 19 February 2015
Team:Sumbawagen/Team
From 2014.igem.org
Notebook – Protocol – 2.PCR (Polymerase Chain Reaction)
1. Ordered primers were dissolved in 1x TE buffer to get a 100 uM stock, then diluted to 10 uM stock by 1x TE buffer (usually to final volume of 200 ul). Primers stocks were stored at -20 °C.
2. There were 3 pairs of primers for the PCR namely
- ACEF/ACER1 to clone catalytic domain sequence of adenylate cyclase gene
- AEF/AER to clone full length sequence of IIA(Glc) gene
3. Reagents compositions for PCR were as follow:
- 2X PCR master mix (Thermoscientific), 25 ul
- Sterilized distilled water (Akuabides), 15 ul
- 10 uM forward primer, 5 ul
- 10 uM reverse primer, 5 ul
- Sterilized distilled water (Akuabides), 15 ul
- 10 uM forward primer, 5 ul
- DNA template, colony from agar plate 0 ul Total volume, 50 ul
4.Reaction conditions for PCR were as follow:
- 95 °C, 2 minutes
- 95 °C, 30 seconds
- 53 °C, 30 seconds
- 72 °C, 2 minutes
- 72 °C, 7 minutes Total cycle, 35 cycles
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