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Unpurifed DNA for methylation - (Aug/08/2005 )

I did some Genomic extraction by using Qiagen Mini Kit, and a lot of samples have the A260/A280 values which are around 1.2. That means my DNA have a lot of contaminants such as RNA and Protein. Does anybody think that would affect the MSP results? Since I am getting weak bands and strong dimers which is not allowed in SYBR real timer PCR.

-zzylx-

yes they certainly affect bisulfite conversion efficency.

is it possible to further purify the DNA with a phenol clhoroform step, precipitate and resuspend for bisulfite conversion.

It is known that protein and RNA contamininants reduce the efficency of the bisulfite conversion reaction.

Nick

-methylnick-