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how to design a probe for unmethylated gene

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#1 nazanin kia

nazanin kia


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Posted 11 January 2013 - 04:21 AM

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

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