Difference between revisions of "Team:EPF Lausanne/Notebook/Yeast"
| Line 39: | Line 39: | ||
<li class="active"><a href="#integrate_pTPGI_dCas9_VP64">Integrate pTPGI_dCas9_VP64</a></li> | <li class="active"><a href="#integrate_pTPGI_dCas9_VP64">Integrate pTPGI_dCas9_VP64</a></li> | ||
<ul> | <ul> | ||
| − | <li | + | <li <a href="#Integrate_reporter_plasmid">Integrate reporter plasmid</a></li> |
<li><a href="#synthesize_promoters">Synthesize promoters</a></li> | <li><a href="#synthesize_promoters">Synthesize promoters</a></li> | ||
<li><a href="#PCRoverlaps_prom">Add Gibson overlaps to CYC promoters by PCR</a></li> | <li><a href="#PCRoverlaps_prom">Add Gibson overlaps to CYC promoters by PCR</a></li> | ||
| Line 45: | Line 45: | ||
<li><a href="#integrate_reporters_plasmids">Integrate the reporter plasmids</a></li> | <li><a href="#integrate_reporters_plasmids">Integrate the reporter plasmids</a></li> | ||
</ul> | </ul> | ||
| − | <li><a href="#Integrate_and_express_gRNAs">Integrate | + | <li><a href="#Integrate_and_express_gRNAs">Integrate and express gRNAs</a></li> |
<ul> | <ul> | ||
<li><a href="#synthesize_gRNAs">Synthesize gRNAs</a></li> | <li><a href="#synthesize_gRNAs">Synthesize gRNAs</a></li> | ||
Revision as of 14:15, 17 August 2015
saccharomyces cerevisiae
pTPGI_dCas9_VP64 integration
As we need dCas9 expressing yeasts, we decided to integrate a plasmid that would inducively express dCas9.
First and second trial
The negative and positive control worked fine, but the transformation control didn’t work. After investigation we found out that we didn’t have an origin of replication in our plasmid thus not enabling our yeasts to replicate the plasmid and to grow. See more details here
