Kun:LabNotes/ASE/2007-10-15
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Exp1: Fixing the PGP1Ver2 PCR reactions I did on 2007-10-12[edit]
- Since I haven't run the thermalcycling on the 96 reactions on cDNAs, I'm just adding the ThermoStart Taq enzyme and more ABI's dNTPs into the reactions.
10x buffer 0.1 x100 10 ThermoStart Taq 0.25 x100 25 10mM dNTP 0.5 x100 50 H2O 0.15 x100 15
I used the aluminum tape to seal the plate. It turned out that I had a evaporation issue. I don't think this plate is useful.
- Also set up 32 reactions on the genomic DNA template.
Reaction set up:
' | one reaction | 33 reactions |
Template | 1 | each |
10x Buffer | 5 | 165 |
25mM MgCl2 | 3 | 100 |
10mM dNTP | 1 | 33 |
10uM primer mix | 1 | each |
50x SYBG I | 0.4 | 13.2 |
ThermoStart Taq | 0.25 | 8.25 |
H2O | 38.35 | 1267 |
95C 15min -> 40 cycles of (94C 30S -> 56C 30S -> 72C 1min) -> 72C 3min, using the PTC-200 machine.