bisulphite chemical modification

Darren A. Natale dnatale at box-d.nih.gov.nospam
Tue Jun 2 09:54:14 EST 1998


Dr. A. Ryan wrote:
> 
> I wish to bisulphite treat a small (10-50ng) sample of DNA for
> X-chromosome inactivation analysis. However, when I follow the methods
> quoted in:
> 
> HERMAN_JG, JEN_J, MERLO_A, BAYLIN_SB CANCER RESEARCH, 1996, Vol.56,
> No.4, pp.722-727,
> 
> I can only recover enough modified DNA if I start off with 10-100 X the
> amount of starting material that I have available. Their methods work
> fine in my hands, but only on lots of good quality DNA, not the piddling
> little amounts of DNA that I have managed to extract from archival
> paraffin sections. So....has anyone come across a small-scale bisulphite
> chemical modification proceedure?

According to Rein et al.(1998) NAR 26(10):2255-2264, you should be able
to
get away with using as little as 10 ng with the bisulfite procedures
referenced
within (it is a review article on methyl-C detection).

D. Natale
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