Difference between revisions of "Team:EPF Lausanne/Notebook/Ecoli"
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| − | <h3> | + | <h3>E. Coli Laboratory Notebook</h3> |
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| + | <a class="dropdown-toggle" data-toggle="dropdown" href="#pdcas9w">Make pdCas9-w<span class="caret"></span></a> | ||
| + | <ul class="dropdown-menu"> | ||
| + | <li><a href="#PCRpdcas9w">PCR</a></li> | ||
| + | <li><a href="#taqpcr">Taq PCR</a></li> | ||
| + | </ul> | ||
| + | </li> | ||
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| − | <!-- | + | <!-- PCR PDCAS9 AND W--> |
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<div id="PCRpdcas9" class="panel"> | <div id="PCRpdcas9" class="panel"> | ||
| − | < | + | <h1>Make pdCas9-w:</br>Open pdCas9 and extract w subunit from pWJ66 by PCR</h2> |
| − | < | + | <p><small>We received plasmids pdCas9 and pWJ66 in bacteria. We did a Miniprep (cf. Protocols) on overnight cultures to isolate them.</small></p> |
| − | < | + | <h2>05.06.2015</h2> |
| − | < | + | <h3>Materials and method</h3> |
| − | + | <ul> | |
| − | + | <li>20 µl Phusion PCR (cf. Protocols) with 1 ng pdCas9 and 1ng of pWJ66, HF buffer and corresponding primers</li> | |
| − | + | <li>PCR product purification (cf. Protocols)</li> | |
| + | <li>Agarose gel electrophoresis of purified PCR products</li> | ||
| + | </ul> | ||
| + | <h3>Results</h3> | ||
| + | <img src="http://https://static.igem.org/mediawiki/2015/e/ee/7.05_pcr_gel.jpg"> | ||
| + | <p><small>Gel shows that extraction of w subunit from pWJ66 was successful, but opening of pdCas9 was not.</br></small></p> | ||
| + | <h2>05.10.2015</h2> | ||
| + | |||
| + | </div> | ||
Revision as of 19:14, 27 July 2015
E. Coli Laboratory Notebook
Make pdCas9-w:Open pdCas9 and extract w subunit from pWJ66 by PCR
We received plasmids pdCas9 and pWJ66 in bacteria. We did a Miniprep (cf. Protocols) on overnight cultures to isolate them.
05.06.2015
Materials and method
- 20 µl Phusion PCR (cf. Protocols) with 1 ng pdCas9 and 1ng of pWJ66, HF buffer and corresponding primers
- PCR product purification (cf. Protocols)
- Agarose gel electrophoresis of purified PCR products
Results
Gel shows that extraction of w subunit from pWJ66 was successful, but opening of pdCas9 was not.