Fig. 3.

Evaluation of a WT-selective PCR blocker by direct sequencing. (A) Plasmids carrying WT sequence or the A1762T/G1764A mutation were mixed at different ratios and served as PCR templates in a SYBR green real-time PCR. The amplification conditions were the same as for Fig. 2 except that the reverse primer was changed to R2 to generate a longer amplicon suitable for sequencing. (B) Sequencing data (for the reverse strand) for nucleotide positions 1765 to 1761. ACCTT represents 1761-AAGGT-1765 (WT), while ATCAT represents 1761-ATGAT-1765 (A1762T/G1764A double mutation).