IGEM:PennState/Labbook/NoahJohnson/2007-8-1

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August 1, 2007

  • Ran gel on PCR products
  • Start @ 11:00

Well

  • 1 . 2-log ladder
  • 2 . 1kb ladder
  • 3 . crp* Taq 75C
  • 4 . (-) Taq 75C
  • 5 . crp* Pfu #1 75C
  • 6 . crp* Pfu #2 75C
  • 7 . crp* Pfu #3 75C
  • 8 . (-) Pfu 75C
  • 9 . crp* Taq 72C
  • 10 . (-) Taq 72C
  • 11 . crp* Pfu #1 72C
  • 12 . crp* Pfu #2 72C
  • 13 . crp* Pfu #3 72C
  • 14 . (-) Pfu 72C

PCR didnt seem to work with Pfu

  • Re-ran large gradient PCR with gradient from 55C-72C
  • Ran Gel from PCR products

Results:

  • no crp* seen
  • probably a problem with Pfu PCR protocol

PCR rxns with Pfu havent been working so I'm going to dilute the primers 10:1 because we think that they could be too concentrated. All other parts of the protocol I will keep constant but I will add 1uL of each primer diluted 10:1 and run a gradient PCR from 61C-72C.

  • Negative ran at 71.4C
  • Ran gel on PCR products (Started @ 7:05pm)

Well:

  • 1
  • 2 . 1kb Ladder
  • 3 . crp* Pfu 72C
  • 4 . crp* Pfu 71.4C
  • 5 . crp* Pfu 68C
  • 6 . crp* Pfu 63.3C
  • 7 . crp* Pfu 61C
  • 8 . (-) Pfu 71.4C
  • 9 . crp* Taq 71.4C


9:30-5:30 and 6:15-7:45

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