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SYBR Green Real time PCR - (Nov/26/2015 )

hello folks,


we are perplexed about a sybr green real time pcr melting curve.

never had problems with this alw, but from one day to the other we observe multiple melting curves - see attachment


We know, that the RNA quality is ok, because all the other PCRs work.


what could this be?

we need help with this!


Thank you in advance


Usually this means you have multiple DNA fragments amplified. Have you run a gel? I predict you'll see two bands. Your primers are probably not sufficiently specific, or there may be duplication of the gene.


Please do not post in multiple fora. The post you made in PCR included a  picture of the melting curve. It appears as if the multiple curves are simply the result of more product in one of the wells than the others. The peaks are at the same temperature. Likely these are the same products (more than one, I think).