IGEM:PennState/Labbook/NoahJohnson/2007-7-31

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July 31, 2007

but with new Pfu polymerase

  • Start @ 12:10
  • Ran gel on PCR products from morning

Samples:

  • 3uL PCR Product
  • 1uL 5x buffer
  • 1uL Sybr Green

Ladder:

  • 1uL ladder
  • 2uL 5x buffer
  • 1uL Sybr Green
  • 6uL dH2O

Well

  • 1 1kb ladder
  • 2 2-log ladder
  • 3 (-) Pfu crp*
  • 4 crp* Pfu #1
  • 5 crp* Pfu #2
  • 6 crp* Pfu #3
  • 7 crp* Pfu #4
  • 8 crp* Pfu #5
  • 9 crp* Pfu #6
  • 10 (-) Taq crp*
  • 11 crp* Taq
  • 12 XylR

PCR wasn't successful for the Pfu crp* runs so ran another PCR with new thermocycler protocol:

  • Cycle 1:(1x)
    • Step 1: 95C for 1:00
  • Cycle 2:(28x)
    • Step 1: 95C for 0:15
    • Step 2: 55C for 0:30
    • Step 3: 72C-75C for 0:45
    • Gradient: H=72C . A=75C
  • Cycle 3:(1x) Hold at 4C


9:30-6

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