Hello,

I plan to perform methylation-sensitive HRM analysis of maize samples.

1. Are methylation-sensitive primer pairs enough or is it necessary to design both, primer pairs for the methylated as well as for the unmethylated version. And in case of methylation-sensitive primers: in plants, this means, that no C of the primer is converted into T. Or is it better to use degenerated primers? What is the best strategy in my case?

2. Most primer design programms are for mammals, I found one for plants, but it is for pyrosequencing not for HRM. Do you know a special programm for my case?

3. Most standards (fully methylated DNA and fully unmethylated DNA) are from human DNA. Where can I find maize standards? How to produce own standards?

Thank you very much!!!