My Notes II

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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
    • Take existing Chris’s {FimH}.{PhoA!} (aka. pBca1102-Bca9194), set up PCR using ca998 oligo and mea37 (<phoA> reverse oligo) to remove stop codon
    • At first, used wrong reverse oligo, so the pcr product came out mysteriously large…until we figured that I was using the wrong oligo.
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