We have been using Thermofisher Melt Doctor for HRM analysis in a Qiagen´s Rotorgene Q. Some results were different from previous experiments with BioRad Precision Melt in a Biomolecular System´s Mic. There is a lot of differences in the cycling and holding conditions among different reagents (Biorad Precision Mix x Melt Doctor x Qiagen Type-it). Any tips on setting up the PCR conditions with this configuration?
Thanks
Hello.
High Resolution Melt (HRM) analysis is a technique that allows for the detection of genetic variations by monitoring the dissociation of double-stranded DNA during PCR amplification. However, the technique can be affected by a number of factors, including the PCR conditions and the reagents used.
When using different reagents and instruments for HRM analysis, it's important to optimize the PCR conditions to ensure that the results are consistent and reliable.
Some tips for setting up the PCR conditions with the configuration you described include:
It's worth noting that these are just general tips, and the specific conditions will vary depending on the samples, primers, and other factors. Therefore, you should perform a systematic optimization process in order to determine the best conditions for your specific application and validate them using the appropriate controls and experiments.
Regards.
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