Sybre green taq man PCR assay for quantitative methylation detection - (Jul/29/2009 )

Dear All,

Im new to taqman assay. Can anyone help me with the probe design and protocol for taqman ? Can i use the MSP primers itself ? i did MSP on my significant CpG islands and i could see the band for methyaltion specific primers, though there is also a light band on unmethylation specific primers (could be due to normal cells in the genomic DNA, no idea ?? or may not be 100 % methyalted)
But i want to quantify methyaltion with a reference DNA, so thought of going for taqman. Any help will be appreciated.

Regards,
RKA

For taqman assay, you need a probe beside the PCR primers. If you do sybr green, no additional probe is required.

pcrman on Aug 1 2009, 08:17 AM said:

Dear Pcrmen,

Can you tell me how much BS converted DNA is advisable to use for sybre green quantitative pcr ?? would be great if you can send any publications with the same method.

Thank you again...Raju