User:Meng Xiao He/Notebook/fall08/2009/01/07

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PCR from yesterday

Phusion runs were with the outer primer set (only the specific pair, no others), Platinum runs were with X over primer set

2009-01-07 pcr.jpg

  • 1st block conditions: Phusion, 12-8 program, unmodified
  • 2nd block conditions: 3% DMSO + Phusion, in gradient thermocycler under 1-6DMSO, seems to work much better
  • 3rd block conditions: Platinum, with Xover primer set


The following from above were rerun:

  • lane 1: cco # 34
  • lanes 20-30: cco (18, 21, 30, 31, 32), cbb (25, 26, 27, 28, 29)

2009-01-07 rerun2.jpg

The lane order is the same as in the bullets, except for the first 3 lanes, which has cco#18 > cco#34 > cco#21

  • lane 1 (cco#18) and lane 6 (cco#31) are the only candidates for loss> there does appear to be banding of some sort > Xover primer set nested PCR candidate
  • lane 5 not explained: did not lose WT copy, and lower band not right size for flanks