Kun:LabNotes/CpgSeq/2007-11-1: Difference between revisions

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(New page: ===Exp. 1 Bisulfite conversion of genomic DNA with EpiTect kit=== Genomic DNA samples (conc. measured by Nanodrop): *DNA RL (from Dr. Huidong Shi), 185ng/ul *Jurkat gDNA (from NEB), 194ng/...)
 
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     DNA Protection Buffer      35ul x 2      35ul 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.
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. Check with argarose gel.

Revision as of 19:05, 2 November 2007

Exp. 1 Bisulfite conversion of genomic DNA with EpiTect kit

Genomic DNA samples (conc. measured by Nanodrop):

  • DNA RL (from Dr. Huidong Shi), 185ng/ul
  • Jurkat gDNA (from NEB), 194ng/ul

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

                                  RL          Jurkat
    DNA                          5ul x 2       5ul x 2
    Water                       15ul x 2      15ul 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. Check with argarose gel.