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Team:The University of Alberta/20 June 2008
From 2008.igem.org
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==Today== | ==Today== | ||
| + | [[Image:june20_purple_russian_5_colonypcr.jpg|thumb|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 [[Team:The_University_of_Alberta/Sequencing|sequencing]] page). Sooo... | *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 [[Team:The_University_of_Alberta/Sequencing|sequencing]] page). Sooo... | ||
**...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. | ||
| - | *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 | + | *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: | ||
| - | Kanayama et al. discusses a E. coli based assay for nuclear receptor ligands [IMPORTANT! or at least useful] | + | --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 | + | --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 | + | --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'''<br> | ||
| + | *Designed primers for the plant primers | ||
| + | *Research into mutation protocols | ||
| + | |||
| + | '''XL1 - Red mannual'''<br> | ||
| + | http://www.stratagene.com/manuals/200129.pdf | ||
Latest revision as of 16:16, 23 June 2008
Today
- 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
