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

Results:
 * Ran Gel from PCR products
 * 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