Help with melt curve
I am very new to qPCR. The problem I face is regarding the melt curve of 1 set of primers.
I performed a trial run and the melt curve gave only 1 peak.
However, the next day I ran the real analysis using the same samples, and the melt curve gave multiple peaks.
This is observed in all replicates from 3 samples. All the other primers are good, even the NTC for this primer is good (no peak at all).
I have ruled out contamination from the reagents as well as the template. I also took cautionary steps to reduce pipetting errors; I had some problems regarding pipetting errors in previous runs. I'm really lost as to what is the cause of this observation. It would be nice if you could suggest some reasons and how could I prevent this from happening again.