User:Meng Xiao He/Notebook/fall08/Constants

=PCR=

cco MR-1 colony
12-7 on big room machine:

25ul reactions, 40 cycles:
 * 7 min initial 94C denaturation
 * 30 s denature
 * 30 s annealing 58→56.5 in first 15 cycles by dropping 0.1 decrease/cycle, 59→56.5 in last 25 cycles
 * 2 min elongation
 * 10 min final extension

Phusion
PhFLANK:
 * 1) 2min at 98
 * 2) 7s at 98
 * 3) 30s @ 63->57 in 15 cycles (drop 0.4 degrees per cycle)
 * 4) 30s @ 59->56 in 15 cycles (drop 0.2 degrees per cycle)
 * 5) 30s @ 57 for 10 cycles
 * 3:45 @ 72 (should capture WT cells as well). 45s for last 10
 * 1) 5 min@ 72 to end
 * 2) hold at 4