multiplex methylation analysis (bs conversion based) - (Jan/20/2010 )
I want to investigate methylation of 10 regions from different CGIs. Does any one have a good idea where i can do a multiplex reaction, so that i only use 10, 20 or 30 ng initial bs converted DNA. I have to work on urine dna assuming the urothelial cells shed off in urine, therfore i have very limited amounts of DNA. This is the reason was thinking about a multiplex reaction. I read about HI-SA technique described by Takeshi Ngasakha in JNCI, but couldnt understand the concept of it. I would be grateful if someone can help me with it....thanks in advance..the paper is attached..the title of the article is "analysis of fecal DNA methylationto detect gastrointestinal neoplasia"....any other ideas are welcome...plzzz help me sooon...
Have you looked into whole genome amplification? That will give you enough DNA for PCR.