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

July 31, 2007

• Ran PCR run identical to one from [July 25, 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