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?