User:University of Washington/7 August 2008

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Contents

Yeast Shuttle Plasmid

- Obtained yeast strain K. lactis

- Obtained primers for amplification of LAC4-LAC12 region of K. lactis genome

- PCR'd the K. lactis with primers

- Made glyercol stocks of BBa_J63005 and BBa_J33202

BioBrick Promoter Measurements

- Inoculated overnight cultures with colony isolates of TOP10 for promoter constructs I20260, I20259, I20261, I20267, and empty TOP10 cells.

LuxR from AraC and TetR(Faifan)

- Both ligated DNA(Elowitz's promoter + pSB1AC3) transformation grew on Amp. Got single colonies. Picked and overnight.

- Continued QuikChange

  • Added 1 ul dpn1 in reaction 1, 2 and negative control, mixed and span 1 min.
  • Incubated 37 degree Celcius for 3 hours
  • PCR purified
  • Transformed 2 ul of each into XL1-Blue, grew on Amp plate

MG1655Z1(Faifan)

-Diluted 1:100 of overnight culture of MG1655Z1(2 colonies from Tsy#2 plate) in Tsy broil.

-Incubated the dilution 3 hours

-Diluted 1:100 again in Tsy, inoculated in Tsy plates.



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