detection of 5-methyl cytosine (bisulfite method)

Onno Myriam Onno.Myriam at univ-rennes1.fr
Mon Oct 24 08:21:28 EST 1994


I ll tried to use an improved method for detection of 5-methyl cytosine by PCR-based genomic sequencing (Biotechniques Vol16 410-417 1994).
This method is based on bisulfite modification of genomic DNA under conditions whereby cytosine is converted to uracil but 5-methyl cytosine remains unaltered.
I have three questions: Is it necessary (possible) to autoclave the bisulfite/hydroquinone solution ? what is exactly the method to alkali denature genomic DNA ? After bisulfite treatment, is it necessery to add NaI before the incubation with the Glassmil
k?

	Any answer is welcome

	Thanks

	
	Myriam

Onno Myriam
Laboratory of Haematology
University of Rennes I
France
e-mail: Onno.Myriam at univ-rennes1.fr



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