User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/15

iGEM 2011

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Goals Get enought HydG for Gibson Assembly. PCR Reagents FinalReact Master Mix 1(PFOR2) Master Mix 2(HydG) H2O 23 70 70 5x Phusion GC Buffer 10 30.5 30.5 dATP 1 3.01 3.01 dCTP 1 3.01 3.01 dTTP 1 3.01 3.01 dGTP 1 3.01 3.01 Primer fwd 5 15.5 15.5 Primer rev 5 15.5 15.5 DNA template 1 3.01 3.01 DMSO 1.5 4.5 4.6 Phusion HF pol 0.5 μL NA NA Total 50 Cycles Gel instruct. Gel Initial 95°C 5 min 30 Rep 98°C 10s 55°C, 60°C, 62°C 30S 72°C 1:45 Hold 72°C 10min 4°C Lane1 Ladder Lane2 Paulina's HydEF2.1 Lane3 Paulina's HydEF2.2 Lane4 Paulina's HydEF2.3 Lane5 Paulina's HydEF1.1 Lane6 Paulina's HydEF1.2 Lane7 Paulina's HydEF1.3 Lane8 HydG 1 Lane9 PFOR2 1 Lane10 HydG 3 Lane11 HydG 2 Lane12 PFOR2 2 Lane13 PFOR2 3