User:University of Washington/21 August 2008

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LuxR from AraC and TetR(Faifan)

- nanodropped the gel purified DNA at 230 nm

ng/ul260/280260/230
BBa_Elowitz2.11.560.03
LuxR5.81.980.06
GFP5.51.410.02
P1010(KAN)4.01.970.02

- ligation

  • mix
dH20(ul)Buffer(ul)P1010-Kan(ul)BBa_Elowitz(ul)GFP(ul)LuxR(ul)T4 ligase(ul)
GFP1.36253.427.22-1>/td>
LuxR1.2825.03.42-7.31>/td>
  • 1 hr incubate at room temp
  • 15 mins denature at 65 degree Celsius
  • 20 mins filtration
  • transformed 2ul DNA into XL1-Blue, inoculated on Kan plate

MG1655Z1(Faifan)

-None of the four cultures grew on Tet, Kan, Cam

-Only culture#3 didn't grow on Amp

  • Streaked #3 to new Tsy plate
  • grew overnight of #3 in Tsy


LuxR from pLac

--I-insert and R-vector parts were isolated from their gel fragments and ligated together.



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