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Hi every one
I'm working on my thesis subject which is analysing methylation status of CDKN2A gene in a special occupational group.I need to perform quantitative MSP and so I decided to design two primers and a probe specific for methylated gene and two primers and a probe for unmethylated gene.
but there is a problem : HOW to design a probe for unmethyated allele???? it can be veryyyyyyyyyy variable!!!! how can i make sure that a probe for unmethylated allele is reliable?
I would appreciate any kind of clues
