1 reply to this topic

[rudi111]

Hi,
I am doing nested PCRs with a first round using BSP and the second round using the methylationspecific primers. This wirks well, however I do not get an visible amplification if I do the MSP without a round of BSP.
Is that normal?
Do I not have to fear PCR-Bias, because the problem in BSP (eg methylation-independent PCR) preferably amplifies the unmethylated allel, if there is one. Could I not get a false negative result for the methylated allel?
What is your experience?

Thanks for your thoughts, Rudi.

[methylnick]

In theory, your BSP primers should not induce bias, it would sound like your starting material is quite minimal and that you are using a bisulphite conversion method that is destructive to the sample.

have you tried doing two rounds of MSP?