I'm doing QRT-PCR, the melting curve looked like this, what can I know from this curve?
That is showing you the relative fluorescence from low to high temperature.
The inflection point on the steepest part of the slope is the melting temp for your PCR product.
You need to change your graph to show the negative derivative of the change in relative fluorescence over temperature to see the "melt peak".
As for what is means, honestly not much for qPCR. The melt peak is the LEAST informative piece of information. It just shows that you made a PCR product & the relative melting temp for that product.
For your analysis, you need the Ct value for each of your samples + controls.
Good luck!
A first derivative curve would be much easier to interpret - the Y-axis would be
-dF/dT.
This would give a curve similar to the attached image. Most analysis software can produce an image of this type; have a look at the software manual.
Vincent Mulholland Sayanta Bera
Katie A S Burnette Thanks a lot for your answers, now I get it!
Hi,
please select the "Calculate Tm" next to the View: Melting curve, and choose the GOI - Color. Now you will get the typical melting curve.
Best
Stefan
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