Kun:LabNotes/CpgSeq/2008-1-2
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Exp. 1 Perform a capturing experiment with more bis-DNA using bisulfite-PCR product as a control[edit]
Pooled PCR product from bisulfite converted Jurkat genomic DNA: 19ng/ul -> 100nM.
JK RL bis-DNA (01/01/08): 0ul 8ul 8ul JK-PCR amplicon: (0.1nM) 1ul 0ul 0ul 10X buffer 1.5ul 1.5ul 1.5ul 10nM CpG-MIP v2 5ul 5ul 5ul H2O 7.5ul 0.5ul 0.5ul
95C 3min->60C 42h-> add 1ul SLN mix (1U/ul stoffel, 0.5U/ul AmpLigase, 200uM dNTP) -> 60C 1h -> 20 cycles of (95C 1min -> 60C 1h) -> 60C 2h -> 37C 1min -> add 1ul of exo mix (Exo I, Exo III, T7 exo), 0.5ul USER enzyme->37c 2 hour -> 94C 5min -> 4C hold.