Difference between revisions of "Team:EPF Lausanne/Notebook/Ecoli"
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</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
| − | <a class="dropdown-toggle" data-toggle="dropdown" href="# | + | <a class="dropdown-toggle" data-toggle="dropdown" href="#">Make pdCas9-w<span class="caret"></span></a> |
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
| − | <li><a href="# | + | <li><a href="#PCRpdcas9">PCR</a></li> |
<li><a href="#taqpcr">Taq PCR</a></li> | <li><a href="#taqpcr">Taq PCR</a></li> | ||
</ul> | </ul> | ||
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</ul> | </ul> | ||
<h3>Results</h3> | <h3>Results</h3> | ||
| − | <img src=" | + | <img src="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> | <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> | <h2>05.10.2015</h2> | ||
Revision as of 19:16, 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.