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DNA quality for MSP? - (Feb/18/2008 )

I am now setting up experiments related with DNA methylation, firstly methylation-specific PCR(MSP).

The first problem that I met is about extracting genomic DNA from mouse tissue, so I found a fast isolation protocol from 'Molecular Cloning', which says the isolated DNA may be about 20-50kb. Maybe some fragmentation occurs according to the protocol.

I thought it would be OK for MSP,
but I'd like to ask you if this protocol is really OK for my experiment.

-awhite0-

The quality in terms of fragmentation is OK. Actually in order to achieve complete bisulfite conversion we sometimes intentionally break up DNA into small pieces by mechanic shearing or enzymatic digestion. In your case the size is perfectly OK, but make sure the DNA is pure, which is also important for complete conversion.

-pcrman-

rolleyes.gif

Thank you so much!!^_^
So, after I purify the DNA, is it enough that I check the purity by measuring the 260/280 ratio?


QUOTE (pcrman @ Feb 18 2008, 08:55 PM)
The quality in terms of fragmentation is OK. Actually in order to achieve complete bisulfite conversion we sometimes intentionally break up DNA into small pieces by mechanic shearing or enzymatic digestion. In your case the size is perfectly OK, but make sure the DNA is pure, which is also important for complete conversion.

-awhite0-