Sodium Bisulfite

muszynski at BIOSCI.MBP.MISSOURI.EDU muszynski at BIOSCI.MBP.MISSOURI.EDU
Tue Jul 6 17:12:14 EST 1993


July 6, 1993

To whoever can help,

I am a post-doctoral fellow at the University of Missouri-Columbia, USA. 

I want to use the bisulfite DNA modification protocol as outlined in the
1992 PNAS paper "A genomic sequencing protocol that yields a positive
display of 5-methylcytosine residues in individual DNA strands."
(89:1827-1831).  I need to look at events represented in small clonal
somatic sectors of tissue so the bisulfite/PCR sequencing is more useful
than using the ligation-mediated genomic sequencing procedure.

But, alas, I am having problems with what should be an easy step.  I'm not
sure how to make up the 3.1 M bisulfite solution.  Earlier parers made
fresh bisulfite soltions at pH 6.0 or 6.1 from a mixture of sodium
bisulfite (NaHSO3) and sodium sulfite (Na2SO3).  The sodium bisulfite
available from all the chemical companies I can find is a mixture of sodium
bisulfite and sodium metabisulfite (Na2S2O5) in some unknown ratio.  Does
this make a difference?  Or should I just assume whatever is in the bottle
labeled "bisulfite" is pure sodium bisulfite?

If anyone has any experience with this protocol or can point me in the
right direction, please email me back as I do not subscribe to the METHODS
newsgroup.  I have emailed my questions to the authors of the above paper
(Peter L. Molloy and Marianne Frommer; CSIRO) but they have not yet
responded.

Thanks for any and all help.

Mike Muszynski
101 Tucker Hall
Univ. of Missouri-Columbia
Columbia,  MO  65211
(314) 882-1168 lab
(314) 882-0123 fax
muszynski at biosci.mbp.missouri.edu




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