User:University of Washington/13 August 2008

Conjugation (Scott)
experiment from Aug. 7 results

LuxR from AraC and TetR(Faifan)
QuikChange Approach

- Overnight QuikChange#4 culture from Amp plate (reaction1 x 4, reaction2 x 2) in Tsy+Amp

- Sequence of DNA from QuikChange#3 came back ==> No mutation.

Elowitz

-Made Glycerol Stock of BBa_Elowitz x 2

-Restriction Digest of BBa_Elowitz, BBa_E0240(GFP), BBa_I0462(LuxR), BBa_P1010(on pSB3K3)
 * mix
 * incubate 37 degree Celsius, 1.5 hours
 * 15 mins water bath 80 degree Celsius
 * nanodrop at 230 nm
 * store in 4 degree Celsius

Lambda Red Recombineering of RP4 (Bryan)
Attempted the following transformations RP1 + pKD78, RP4 + pKD78, and pKD78 alone. Selected with cam/tet, cam/tet, and cam, resepectively. None of the O.N. cultures grew. Possible problems are imprecision in micropipetting due to extremely small volumes <0.5 ul of vector DNA or not enough cells (1 mL of culture per electroporation).

AHL Expression in Yeast (Bryan)
Transformed C0161/YEp426 ligation produce from yeseterday into XL-1 Blue. O.N. culture grew in amp, indicating successful transformation. Miniprepped and made glycerol stock.

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