Kun:LabNotes/CpgSeq/2008-1-1

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Exp.1 Bisulfite conversion of DNA[edit]

Resuspend one vial of Bisulfite Mix in 800ul nuclease free water, votex for 5min. Set up reactions:

                                  RL          Jurkat
    DNA                         10ul x 2      20ul x 2
    Water                       10ul x 2      
    Bisulfite mix               85ul x 2      85ul x 2
    DNA Protection Buffer       35ul x 2      35ul x 2

Perform the bisulfite conversion on PTC-200 using the EpiTect program 99C5min->60C20min->99C5min->60C1h25min->99C5min->60C2h55min->20Chold.

Follow the EpiTect protocol to clean up the bisulphite converted DNAs.

Concentrate to 5ul with Speedvac.