Calculated ΔTm dependency on nucleotide mutation and position in 20mer oligonucleotides. (A) Average ΔTm values of all sequence changes at each position are shown with standard deviations. (B) Average ΔTm values of each sequence change are shown. Data point colors represent the original sequences: blue, A; pink, T; green, C; and orange, G, while shapes represent the mutated sequences: square, A; diamond, T; circle, C; and triangle, G (blue diamonds, from A to T; blue circles, A to C; blue triangles, A to G; pink squares, T to A; pink circles, T to C; pink triangles, T to G; green squares, C to A; green diamonds, C to T; green triangles, C to G; orange squares, G to A; orange diamonds, G to T; and orange circles, G to C). (C) ΔTm dependency on nucleotide in 20mer oligonucleotides excluding the three end points. (D) All ΔTd values of each sequence change from Urakawa et al. () are shown at each position from the 5′-end excluding the three end positions. Data from oligo 1 and oligo 2 in Figure are not distinguished. Color and shape representations are as in (B). A data point indicated with an arrow is a [G, C] mismatch case, which will be ignored in the comparison.