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User:University of Washington/8 September 2008
From 2008.igem.org
LuxR from AraC and TetR(Faifan)
-Sequences of LuxR construct came back. 4/8 contained P1010, the other 4/8 had too many N's-cannot determine.
-Screening 14 colonies by colony PCR
- resuspended each colony into 50 ul dH2O, vortexed
- mix (10 ul rxn)
- 5.95 ul dH2O
- 1 ul 10x Taq Buffer
- 0.25 ul 10mM dNTPs
- 0.50 ul MgCl2
- 0.6 ul VF2(10 pmol/ul)
- 0.6 ul VR(10 pmol/ul)
- 0.1 ul Taq
- 1 ul colony suspension
- Thermocycling
- 1 cycle : 95 C : 6 mins
- 35 cycle: 95 C : 30 s
- ________: 53 C : 30 s
- ________: 72 C : 2 mins
- 1 cycle : 72 C : 10 mins
- 1 cycle : 4 C : forever
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