Team:The University of Alberta/20 June 2008

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**...She is doing colony PCR on colonies taken from the streaked-out "Colony 5" plate using both the V<sub>F</sub>/V<sub>R</sub> primers and the gene-specific P<sub>F</sub>/P<sub>R2</sub> 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 ]).
**...She is doing colony PCR on colonies taken from the streaked-out "Colony 5" plate using both the V<sub>F</sub>/V<sub>R</sub> primers and the gene-specific P<sub>F</sub>/P<sub>R2</sub> 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.
**...sequence the correct band so we can be doubly sure that the "Purple Russian Colony #5" plate really is what we think it is.
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*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 '''COMING SOON''').
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*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:
*Mike posted links (on Project page) to papers discussed in yesterday's meetings, and to sequences that have been ordered:
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--Zuo et al. discusses a fusion protein that uses LBD of ER
--Zuo et al. discusses a fusion protein that uses LBD of ER
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* We have also opted to use a mutator strain deficient in repair mechanisms to induce purple russian and get purple expression
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* 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'''<br>
'''Jason'''<br>
*Designed primers for the plant primers
*Designed primers for the plant primers
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* Research into mutation protocols
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*Research into mutation protocols
'''XL1 - Red mannual'''<br>
'''XL1 - Red mannual'''<br>
http://www.stratagene.com/manuals/200129.pdf
http://www.stratagene.com/manuals/200129.pdf

Latest revision as of 16:16, 23 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 VF/VR primers and the gene-specific PF/PR2 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

  • Designed primers for the plant primers
  • Research into mutation protocols

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

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