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

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