I observe double peaks in RT-PCR for the primers that were validated using gel electrophoresis in other samples. I have used these primers on three different sample sets (about twenty-four samples in each) and the data was clean.

In this case, I am using sixteen samples and these primer dimers observed (ran it on a gel) were not consistently found in all of these sixteen samples. I tried for two other genes, these primers gave clean results for all the data sets including the other three data sets. Now I am really confused - because this clearly does not seem to be neither a problem of primers nor of the sample quality. Why are these primer-dimers observed selectively only in some samples? I am not sure whether to proceed with the data I got from the RT run.

Can anyone help me with this issue ?

More Srividya Vaidyanathan's questions See All
Similar questions and discussions