Difference between revisions of "Team:EPF Lausanne/Notebook/Ecoli"
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<a class="dropdown-toggle" data-toggle="dropdown" href="#">Make pdCas9-w<span class="caret"></span></a> | <a class="dropdown-toggle" data-toggle="dropdown" href="#">Make pdCas9-w<span class="caret"></span></a> | ||
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.