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

July 31, 2007

but with new Pfu polymerase
 * Ran PCR run identical to one from [July 25, 2007]
 * Start @ 12:10

Samples:
 * Ran gel on PCR products from morning
 * 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