How can I calculate methylation level from melting curve or melting peaks from Light Cycler 480 qPCR?

20 December 2016 1 4K Report

I am trying to quantify methylation levels for my DNA samples with high resolution melting (MS-HRM). I have done melting curve analysis with my samples and standards (100%, 75%, 50%, 25% and 0%) using LightCycler 480 qPCR-machine and now I have the raw melting curve data. I have tried to analyze the results with Tm calling module and GeneScanning software but I dont know how to calculate the exact methylation levels for my samples from the results. Have anyone done MS-HRM with LightCycler 480 and could tell how they´ve done the calculations?

Rakesh Kumar Sinha

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2679043/

go through the above publication.

It will help you

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