I am having problems with my qPCR. I am trying to amplify the humand ADH1A gene. I have primers (do not know the sequence) that seem to work fine amplyfing the gene and they give me a nice melting curve. However, my negative control (primers only) comes up positive:-( I have repeated the experiment so many times now it is really frustrating. I have been really careful handeling the plate, samples, and reagents. I have used new water and primers.
Could it be that the ADH1A gene is structurally difficult to amplify??
I really appreciate any comments.
Thanks for your help.