Kun:LabNotes/ASE/2007-11-3
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Exp. 1 Perform PCR on the eSNP84k set 1-4 using iTaq.[edit]
template: 0.2ul 1.6ul x 4 x 2 10x buffer: 10ul 80ul x 4 x 2 10mM dNTP: 2ul 16ul x 4 x 2 100uMAP1V41U: 0.4ul 3.2ul x 4 x 2 100uMphosAP2V4: 0.4ul 3.2ul x 4 x 2 Taq: 0.5 or 1ul 8ul x 4 x 2 50x SYBR I: 0.8ul 6.4ul x 4 x 2 H2O: 86ul 680ul x 4 x 2
I set up 800ul reactions for each library first, but then wasn't quite confident that I add the round amount of enzyme and primers to each tube, so decided to do another 800ul.
94C 3min -> 17 cycles of (94C 45sec -> 58C 2min -> 72C 1min) -> 72C 3min -> 15C hold.