I am currently working on HRM analysis of individual CpG sites. I am trying to understand the extent of methylation (%) in cases against controls. However, the derivative melt curve analysis is not giving satisfying results. Can some one please suggest how to get better bell shaped curve? Also, few samples are going beyond 100%.
I am using standards 0% and 100% in equimolar concentrations. (Qiagen make)
The idea that occurs to me is
that it uses in its cases (controls and study samples)
an integrated quadratic model with a logarithmic analysis.
The best thing is that you create a program in the simplest
programming language for biological study (Phyton)
according to your problem.
This will allow you to roughly model the curve you are looking for. I believe that because the data should not
be interpreted in the correct way (lnx) when x is its standard value, I believe that its curve would not exceed (%) what was expected. I hope I have helped you according to my little experience in the subject, it is the answer that I can offer you.
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