Kun:LabNotes/ASE/2007-11-1
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Exp1: Repeat PCR amplification on the eSNP84k library using NEB 2X Taq master mix[edit]
Reactions eSNP84k set1-5 (100nM):
x 8 x 5 Template 0.2ul 1.6ul x 5 2x Master mix 50ul 400ul x 5 100uM AP1V41U 0.4ul 3.2ul x 5 100uM phosAp2V4 0.4ul 3.2ul x 5 H2O 49ul 386ul x 5 50X SYBR I 0.8ul 6.4ul x 5
94C 2min -> 16 cycles of (94C 45sec -> 58C 2min -> 72C 1min) -> 72C 3min -> 4C hold
I added 2 more cycles for #1 and 3 more for #5 since their amplification curves came up a little bit later than others.
Ethanol precipitate the amplicons, 400ul per tube.