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qPCR melting curve trouble

qPCR melting curve trouble - PCR - Polymerase Chain Reaction Forum

qPCR melting curve trouble - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 07-09-2009, 01:31 AM
Pipette Filler
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Talking qPCR melting curve trouble



Dear everyone

I'm new for qPCR only several monthes. And using the PCR production as SYBR Green qPCR's template to generate the standard curve. Normally we use the size below 300bp, but some of my productions have to be around 550pb, is it acceptable? And there are two peaks of the melting curve that quite near to each other. How can deal with it? (I already modified the annealling temperature and primer concentration but useless. )
And another melting curve is very broad but single peak, I'm afraid there are two peaks inside. So maybe it should with the same problem as before?

Sorry for poor English.
Would you help me please?

Last edited by amoon_sasa; 07-13-2009 at 05:49 AM.
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Old 07-17-2009, 09:06 AM
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Default Re: qPCR melting curve trouble

Can you attach the melting curve for us to see? thanks
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