I am doing qPCR for a long a time and trying to optimize conditions for my gene of interest. However, I am encountering abnormal amplification curves that affects the analysis. I am attaching the image.
What could be the reason? Any idea?
Thanks a lot..
Abnormal amplification curve
1 reply to this topic
Posted 03 October 2011 - 08:01 AM
The wave pattern suggests that you have two products -- possibly an off target priming by only one of the primers that eventually goes exponential after it generates enough template (ie acts as both fwd and rev primers to the amplicon)