I am running HRM these days. One thing I never understood is the relation between the length of amplicon and melting temperature. I mean, will temperature increase if length is longer? I am guessing that the answer is negative. Probably melting temp does not work that way.
I have a set of primers that amplify a 120 bp amplicon. Then I have another set that amplify a 300 of the same gene. How different their melt peak should be? Definitely, it does not mean that the 300 bp amplicon must show a higher melting peak than that of the 120 bp? Correct? If the 120 bp product shows a peak at 81 C, it does not mean that the 300 bp product must be above 81 C? I have problem understanding this.