User:University of Washington/5 September 2008

From 2008.igem.org

LuxR from AraC and TetR(Faifan)

- measured Absorbance of the overnight culture at 660 nm = 1.5

- diluted 60 ul culture into 5.40 ml Tsy+KAN. Absorbance at 660 nm = 0.035

- incubated 37 degree Celsius for 2.5 hrs, Absorbance at 600 nm = 0.7

- diluted 1 ml culture into 5 ml Tsy+Kan, OD@600nm = 0.165

- set up 96 well plate reader experiment:

  • 5x5, 160 ul culture
Arabinose(%), 10%Arabinose added (ul)
0%, 0 ul0.05%, 1ul0.1%, 2ul0.50%, 10ul1.00%, 20ul
aTc(ng/ml), 10ug/ml-aTc added(ul)0, 0ulA1A2A3A4A5
50, 1ulB1B2B3B4B5
100, 2ulC1C2C3C4C5
500, 10ulD1D2D3D4D5
1000, 20 ulE1E2E3E4E5
  • added dH2O to final the volume to 200 ul
  • added 40 ul of mineral oil to each well to protect evaporation
  • set the machine to run 24 hrs, read every 10 mins

Conjugation (Scott)

Conjugation method #2
·pAC99 + pUB307

Glycerol Stock
·pAC99
·pAC99 + pUB307


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