User:Meng Xiao He/Notebook/fall08/2009/01/07: Difference between revisions
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==PCR from yesterday== | ==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 | |||
[[Image:2009-01-07_pcr.jpg]] | [[Image:2009-01-07_pcr.jpg]] | ||
*1st block conditions: 12-8 program, unmodified | *1st block conditions: Phusion, 12-8 program, unmodified | ||
*2nd block conditions: | *2nd block conditions: 3% DMSO + Phusion, in gradient thermocycler under '''1-6DMSO''', seems to work much better | ||
*3rd block conditions: Platinum | *3rd block conditions: Platinum, with Xover primer set | ||
===Rerun=== | ===Rerun=== | ||
The following from above were rerun: | The following from above were rerun: | ||
*1: cco # 34 | *lane 1: cco # 34 | ||
*20-30: cco (18, 21, 30, 31, 32), cbb (25, 26, 27, 28, 29) | *lanes 20-30: cco (18, 21, 30, 31, 32), cbb (25, 26, 27, 28, 29) | ||
[[Image:2009-01-07_rerun2.jpg]] | [[Image:2009-01-07_rerun2.jpg]] | ||
The order is the same as in the bullets, except for the first 3 lanes, which has cco#18 > cco#34 > cco#21 | 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 | |||
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Latest revision as of 10:36, 8 January 2009
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PCR from yesterdayPhusion runs were with the outer primer set (only the specific pair, no others), Platinum runs were with X over primer set
RerunThe following from above were rerun:
The lane order is the same as in the bullets, except for the first 3 lanes, which has cco#18 > cco#34 > cco#21
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