User:University of Washington/9 September 2008

From 2008.igem.org

LuxR from AraC and TetR(Faifan)

-Ran gel of the PCR products x14 colonies, got no DNA in all lanes-probably because cells weren't lysed or there weren't any cells at all from the beginning(resuspension wasn't good enough to disperse the cells in dH2O)

-Resuspended 28 colonies into 20 ul dH2O (14 from yesterday's resuspension, 14 from plate), boiled at 100 degree Celsius for 16 mins.

-Did Colony PCR again

  • mix 12.6ul dH2O + 2 Buffer + 0.4ul 10mM-dNTP + 0.8ul 50mM-MgCl2 + 1ul 10mM-VF2 + 1ul 10mM-VR + 0.2ul Taq + 2ul Template
  • Thermocycling
    • 1 : 95 : 1 min
    • 30: 95 : 30 s
    • __: 53 : 30 s
    • __: 72 : 1 min
    • 1 : 72 : 7 mins
    • 1 : 4 : forever

-Grew overnight of MG1655Z1 from stock in Tsy

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