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

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iGEM 2011 Main project page
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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



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