IGEM:PennState/Labbook/NoahJohnson/2007-7-31: Difference between revisions
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Well | Well | ||
1 1kb ladder | *1 1kb ladder | ||
2 2-log ladder | *2 2-log ladder | ||
3 (-) Pfu crp* | *3 (-) Pfu crp* | ||
4 crp* Pfu #1 | *4 crp* Pfu #1 | ||
5 crp* Pfu #2 | *5 crp* Pfu #2 | ||
6 crp* Pfu #3 | *6 crp* Pfu #3 | ||
7 crp* Pfu #4 | *7 crp* Pfu #4 | ||
8 crp* Pfu #5 | *8 crp* Pfu #5 | ||
9 crp* Pfu #6 | *9 crp* Pfu #6 | ||
10 (-) Taq crp* | *10 (-) Taq crp* | ||
11 crp* Taq | *11 crp* Taq | ||
12 XylR | *12 XylR | ||
PCR wasn't successful for the Pfu crp* runs so ran another PCR with new thermocycler protocol: | PCR wasn't successful for the Pfu crp* runs so ran another PCR with new thermocycler protocol: | ||
Latest revision as of 08:05, 1 August 2007
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
