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Real time PCR melting curve - (Dec/03/2012 )

Actually, i face a problem with real time PCR of miRNA. I tried the primer of one type of miRNA two time and i observed a one typical bell shape curve and other multi peak curve . Could any one tell me about the causes of multi peak melting curve in real time PCR

-Mohamed 1984-

Mohamed 1984 on Tue Dec 4 01:31:40 2012 said:


Actually, i face a problem with real time PCR of miRNA. I tried the primer of one type of miRNA two time and i observed a one typical bell shape curve and other multi peak curve . Could any one tell me about the causes of multi peak melting curve in real time PCR

multiple peaks mean multiple products which can be non-specific amplicons or usually from primer dimers but I'm not sure about miRNA primers- are they from a publication?...(posting an image will be more helpful) and what about your NTCs (no peaks?) You can also search for threads in this forum addressing this same or related issues....

-casandra-

Actually, I ordered the miRNA primers directly from company. I do not formulate it.

-Mohamed 1984-