Figure 1.

Detection of FLT3 ITD by high-resolution melting analysis. A: Fluorescence (F) versus temperature (T) melting curves using raw fluorescence data. Four wild-type (black, brown, green, and purple line), and four ITD mutant (red, orange, blue, and light blue line) patient samples are shown. The insertions within the ITD mutant samples ranged from 6 to 102 bp. B: Melting curves after fluorescence-normalization, for the samples depicted in A. The wild-type samples have narrow melting transitions, whereas the ITD mutant samples have broad or multiple melting transitions. C: Temperature shifted melting curves. ITD mutant samples are easily distinguished by the low temperature decrease in fluorescence. D: Fluorescence difference plots of the normalized data. A wild-type sample is used for the baseline and the curves for the remaining wild-type samples cluster around the baseline. The curves for the mutant samples are depicted in the positive scale with a greater fluorescence difference. E: Derivative (-dF/dT versus T) melting curves of the normalized fluorescence data. The wild-type sample has a single peak, whereas the ITD mutant samples have either a low temperature shoulder or two peaks.