methylcytosine mapping

NH at LGC.CO.UK NH at LGC.CO.UK
Tue Mar 3 06:28:22 EST 1998


Dear All,

I'm currently trying to 5-methylcytosine map mammalian genomic DNA using
the bisulfite method. Unfortunately, I'm failing to get specific amplification of
a PCR product using primers designed to the modified sequence (i.e. post
bisulfite treatment, with cytosines changed to uracils/thymine-avoiding
sequences that could be potentially methylated and therefore not change).
I have one paper that suggests certain methods cause template
degradation. I would be very grateful if anyone has any tips/protocols for
5-methylcytosine mapping of genomic DNA from mammalian tissues, that
work in their hands.

I am not a subscriber to this newsgroup so could any electronic replies be
sent to my email address please.

Many thanks,

Dr. Neil Harris,
Analytical Molecular Biology,
LGC,
Queens Road,
Teddington,
Middlesex.
TW11 OLY

Tel: 0181 943 7444
Fax: 0181 943 2767
email: nh at lgc.co.uk




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