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Real time PCR melting curve

Started by Mohamed 1984, Dec 03 2012 05:31 PM

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#1 Mohamed 1984

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Posted 03 December 2012 - 05:31 PM

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

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#2 casandra

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Posted 03 December 2012 - 08:52 PM

Mohamed 1984, on 03 December 2012 - 05:31 PM, 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....

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