Jie:LabNotes/CpgSeq/2009-8-27
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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, MATN4 and DNMT3B at 58C, rs10846023 on 62C 1min, 72C 1min)-> 72C 5min.