Can bisulphite treatment be used for detection of DNA methylation within bacteri - (Oct/22/2007 )
As far as I understand, using bisulphite to treat cells for detection of DNA methylation relies on the methylation of cytosine residues within CpG sites. However, bacterial cells seem to primarily methylate Adenine (within GATC sites). Can anybody let me know whether this means that I will not be able to detect methylation within bacterial cells through bisulphite treatment? And if not, can anybody suggest any different methods that can be used to detect DNA methylation within bacteria?
-Mike-
-Mikelus-
QUOTE (Mikelus @ Oct 22 2007, 08:34 AM)
As far as I understand, using bisulphite to treat cells for detection of DNA methylation relies on the methylation of cytosine residues within CpG sites. However, bacterial cells seem to primarily methylate Adenine (within GATC sites). Can anybody let me know whether this means that I will not be able to detect methylation within bacterial cells through bisulphite treatment? And if not, can anybody suggest any different methods that can be used to detect DNA methylation within bacteria?
-Mike-
-Mike-
Methylation occurs on both cytosine and adenine in Bacteria.
if you can't use BiS treatment for your bacteria, there are other methods.
Look for a restriction enzyme that cleaves only at methylated sites(McrBC, etc). Design an PCR amplicon containing methylated sites. Digest your DNA, and see if you get amplification.. Real time, or end point. It's not at pretty as BiS, but it works.
-sneth-
Many thanks 
-Mikelus-