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.