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My Notes II
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| - | *Decided to shrink old tree and use new methods to finish the tags | + | *Decided to shrink old tree and use new methods to finish the tags: attach my {<tag!}.{b1006} parts to {FimH}.{phoA!} |
| - | **Take existing | + | **Take existing {FimH}.{PhoA!} (aka. pBca1102-Bca9194), set up PCR using ca998 oligo and mea37 (<phoA> reverse oligo) to remove stop codon |
| - | **At first, | + | **At first, the PCR product came out mysteriously large. We tried 3 times and the same thing occurred. |
| + | It was not until 11:00pm, that figured that I was using the wrong oligo ... we set up the 4th PCR then - with the ''correct'' oligo. | ||
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| + | *also amplified Bca9194 for personal stock, since we were running low - grew on amp plate AND amp media | ||
Revision as of 06:11, 11 August 2008
07 August 2008 (Th)
- Picked colonies for sca2-7; streaked on CK plate, screened on Amp. Growing overnight in CK media
- mini meeting - new methods enforced due to assembly status:
Assembly Status:
All transfers completed successfully, EXCEPT sca2 (rbs1-A) and sca3 (fimH)
Layer one assemblies completed:
sca21-{<phoA!}.{b1006}
sca22-{<AP!}.{b1006}
sca23-{<HA!}.{b1006}
sca24-{<myc!}.{b1006}
sca25-{<FLAG!}.{b1006}
None of Layer 2 or 3.
- Decided to shrink old tree and use new methods to finish the tags: attach my {<tag!}.{b1006} parts to {FimH}.{phoA!}
- Take existing {FimH}.{PhoA!} (aka. pBca1102-Bca9194), set up PCR using ca998 oligo and mea37 (<phoA> reverse oligo) to remove stop codon
- At first, the PCR product came out mysteriously large. We tried 3 times and the same thing occurred.
It was not until 11:00pm, that figured that I was using the wrong oligo ... we set up the 4th PCR then - with the correct oligo.
- also amplified Bca9194 for personal stock, since we were running low - grew on amp plate AND amp media
