Fig 3. Differential calculus analysis of HRM data.

A) Representative results of differential calculus analyses of HRM for IDH1^R132. First row: Sanger sequencing; second row: fluorescence intensity curve; third row: -d¹ curve; fourth row: -d² curve. First column: A result of sequence-wild type DNA containing no heteroduplex-derived peaks in either the -d¹ or -d² curves. Second column: A result of sequence-mutant DNA. Whereas the heteroduplex-derived peak is recognized in the -d¹ curve as a slight change of shape, the -d² curve demonstrates a more distinct peak formation. B) Distributions of run-to-run variability in the low-temperature melting transition (LTMT) (blue) and high-temperature melting transition (HTMT) (red) positions from Tm. Bars indicate the number of observations within the bins of width 0.01°C. The curved black line shows the approximate normal distributions of LTMT and HTMT, in which the standard deviations were 0.076°C and 0.048°C, respectively.