Principle of Probe-Based Melting Curve Analysis
Probe-based melting curve analysis is based on melting temperature generated by thermal denaturation of
the probe-target hybrid. Melting curve plot of fluorescence (F) versus temperature (T) is transformed into
melting peak by plotting -dF/dT versus temperature. The hybridization probe with fully matched wild-type
target gives the highest T m value. With matched mutant-type target, it gives the lower T m value.
Mutation is detected as T m deviation (△T m ) compared to the wild-type hybrid.