using restriction enzymes for studying DNA methylation - hemimethylation (Jan/23/2008 )

Hello

How do I identify if a region is hemimethylated (i.e. one allele being methylated and the other unmethylated) when using restriction enzymes HpaII and MspI? I know that MspI cleaves such regions, but does HpaII?

Answer such questions by looking at Rebase, http://rebase.neb.com and looking up the enzyme. The enzyme page has a link to the methylation sensitivity tests, and vast amounts of other information.

Hi there,

MspI will cleave irrespective of Methylation,

HpaII will cleave the same sequences if it is unmethylated, so for an imprinted region there will be partial digestion of the site.

Nick

Thanks!

I had heard somewhere that HpaII does not cleave, even if one of the strands is unmethylated, but the fact then is, that it will cleave, but, so to say, the amplification product (if you were to amplify the HpaII fragments) would be less than if a sequence is totally unmethylated in both strands...