Kun:LabNotes/ASE/2007-11-2
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Exp. 1 Perform PCR on the eSNP84k set using different Tag polymerases.[edit]
- ABI Platinium Taq Gold
- Thermo Start Taq
- Supernova HotStart
- Qiagen HotStar Plus
- Bio-Rad iTaq
- Sigma JumpStart Taq
Template: eSNP84k set #5.
I use the same JumpStart 10X buffer.
template: 0.2ul 1.6ul x 6 10x buffer: 10ul 80ul x 6 10mM dNTP: 2ul 16ul x 6 100uMAP1V41U: 0.4ul 3.2ul x 6 100uMphosAP2V4: 0.4ul 3.2ul x 6 Taq: 0.5 or 1ul 8 or 16ul x 6 50x SYBR I: 0.8ul 6.4ul x 6 H2O: 86ul 680ul x 6
95C 12min (or 5min) -> 94C 3min -> 17 cycles of (94C 45sec -> 58C 2min -> 72C 1min) -> 72C 3min -> 15C hold.
Surprisingly, all reactions looked good based on amplification curves. I added two more cycles since the curves have not reached to plateau yet.