Ran a qPCR using StepOnePlus to test out some new GPER1 primers. The attached images are both for GPER, but are different primers. Has anyone seen a melting curve that looks like this? I cannot determine if the early spikes are from nonspecific binding or primer dimers. My GAPDH curve also has a small spike.
We usually do see a slight first peak in the melting curves but never as high as on the second image. I would assume that these are primer dimers, since their melting temperature seems to be too low for a decent long product. Maybe the PCR runs inefficient and primer dimers can form easier than in the first qPCR using different primers
However 65° seems to be rather high. Do you have calculated melting temperatures for your second primer pair?
Hello ... do you find a solution for your problem? Im having a similar one
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