How to set melting curve program ? Should we use the same temperature annealing of the cycling stage?
Gordon L Warren
Here is a blurb I stole from the Qiagen website. It should answer your question.
"First, all the components are denatured at 95°C, followed by complete annealing at a set temperature (based on the primer Tm values), followed by a gradual increase in temperature up to 95°C. Fluorescence intensity is monitored during this final temperature increase, resulting in the generation of a melting curve or dissociation curve."
During the ramp up to 95°C, I have generally increased by 2°C/s.
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