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-