A question about MSP - (Sep/10/2008 )

Hi, I'm a newer here. Now I meet a problem about MSP.
For my negetive control DNA (unmethylated), I have bands of both U and M primers after modification and PCR, which means I cannot modified my DNA completely. I reduced quantity of DNA and increased temperature of incubation, but have no result. Have you met this problem before? And what can I do to solve it?

Hoping for your answer.Thanks a lot

-biox-

I do not think the modification does not work, if you used methylation kit for treatment. It seems that the primers you are using have problems.

-microlight-

QUOTE (microlight @ Sep 10 2008, 11:18 PM)

I do not think the modification does not work, if you used methylation kit for treatment. It seems that the primers you are using have problems.

Thank you for your help~ but I don't think the problem is the primers, because I use the primers in published articles

-biox-

QUOTE (biox @ Sep 10 2008, 09:34 AM)

Have you met this problem before? And what can I do to solve it?

Hoping for your answer.Thanks a lot

Hi!
Does it happen with all the primers you use?? I mean, maybe the genes you're studing are methylated and you observe both bands (unmethylated and methylated) because of the heterogenity of your cell population... Or maybe you should perform your PCR en a higher rectrictive conditions.

-yarince-

okay, try optimize your PCR conditions. When you still have trouble with your assay, let me know, and show me your primer sequence.

-microlight-