The purple line is the problem. There the curves start raising slowly already at round one. We see this happening very often. Although some wells seems to have this problem more then other wells, it occurs still randomly. Also there is no clear correlation with the primer sequence. With the same primer set, some wells may have the problem while other wells don't have it. We perform the qPCR in triplicate. In general, just one out of these three hows the problem.
Usually there are only a few wells that show the problem, but sometimes more then 80% of the wells show the problem. And in rare cases, all the wells from one primer set shows this problem.
Who can explain what is happening here?
Thanks for your reply.
Edited by theo22, 30 March 2011 - 01:56 AM.