Jie:LabNotes/CpgSeq/2009-8-27: Difference between revisions
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>Sam Chiang No edit summary |
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==TA cloning of 7 samples== | ==TA cloning of 7 samples== | ||
I did TA cloning for | I did TA cloning for | ||
chr12: | chr12:4541690_rs34206253 on PGP1L and PGP1F; | ||
chr21: | chr21:32746340_rs8128908 on PGP1L and PGP1F; | ||
chr20: | chr20:60061824_rs6061990 on PGP3L and PGP9L; | ||
chr20:31718894_rs3746459 on BJ; | chr20:31718894_rs3746459 on BJ; |
Revision as of 15:55, 29 August 2009
ASM validation of cell type and individual dependent ASM
primers for cell and individual dependent ASM validation
I did the PCR using 2x iQ supermix on: chr12:4541690 on PGP1L and PGP1F; chr21:32746340 on PGP1L and PGP1F; chr20:60061824 on PGP3L and PGP9L; chr20:60403281 on PGP1L and PGP3L; chr20:31718894_rs3746459 on BJ;
x 9 2x iQ supermix 50 450 Primer F (50uM) 0.5 0.5 each Primer R (50uM) 0.5 0.5 each sample bis-gDNA (~202ng/ul) 0.1 0.1 each dH2O 50 450 total 40ul
96C 3min-> 45x (95C 30sec, 62C 1min, 72C 1min)-> 72C 5min.
I get the amplicons from all 9 samples except chr20:60403281 on PGP1L and PGP3L.
TA cloning of 7 samples
I did TA cloning for chr12:4541690_rs34206253 on PGP1L and PGP1F; chr21:32746340_rs8128908 on PGP1L and PGP1F; chr20:60061824_rs6061990 on PGP3L and PGP9L; chr20:31718894_rs3746459 on BJ;