Melting temperature of the DNA and HRM

[amicia]

Hi Lazyphant,
In my opinion, low level of amplicons, due either by low amount of start DNA or low efficiency, could interfere cause the systems are not able to detect specific strong fluorescence signals and High Melting Resolution analysis (HRM) could be at lower resolution and the results can vary (due to artifacts or aberrant amplification). This is of particular relevance because the HRM is done comparing different samples. Personally, I think this the main reason why an optimized PCR reaction is essential before a melting curve analysis is done.
Referring to and how the DNA amount and quality could interfere during HMR analysis try to read this: [Only registered users see links. ].