Team:The University of Alberta/20 June 2008

Today

Colony PCR from Purple Russian Colony #5 plate

Got the sequencing results back from the "Purple Russian Colony #5" sequencing that Winnie did. Looks like it didn't work at all (see the sequencing page). Sooo...
- ...She is doing colony PCR on colonies taken from the streaked-out "Colony 5" plate using both the V_F/V_R primers and the gene-specific P_F/P_R2 primers; the former will show us the size of whatever insert is in the colonies, the latter will show a band if and only if the insert is Purple Russian (though this is really redundant, since it is Purple Russian beyond a reasonable doubt [ See colony PCR results below ]).
- ...sequence the correct band so we can be doubly sure that the "Purple Russian Colony #5" plate really is what we think it is.
Last night the volunteers ran the colony PCR Jason did on Purple Russian Colony #5. The band was the right size (~700bp) so we are reasonably sure that this is what we want. (See image to the right).
Mike posted links (on Project page) to papers discussed in yesterday's meetings, and to sequences that have been ordered:

--Kanayama et al. discusses a E. coli based assay for nuclear receptor ligands [IMPORTANT! or at least useful]

--Zhang et al. discusses difficulty of expressing full length ER in E. coli

--Zuo et al. discusses a fusion protein that uses LBD of ER

We have also opted to use a mutator strain deficient in repair mechanisms to try and mutate purple russian in such a way that we will get purple expression.

Jason

XL1 - Red mannual
http://www.stratagene.com/manuals/200129.pdf