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.