FIG. 2.
The effect of sequence mismatches on PCR amplification in real-time PCR assays. Serial dilutions of six different plasmids were performed to generate standard curves from 2.5 × 107 copies to 25 copies. Real-time PCR assays were performed as follows: top, AllBac assay with 0 sequence mismatches to all plasmids; middle, HuBac assay with 0 to 7 sequence mismatches; and bottom, BoBac assay with 0 to 14 sequence mismatches. Base pair mismatches between each plasmid and the primer and probe used in the real-time PCR assay are in parentheses in the legend boxes. Plasmids having more than seven mismatches to the primers and probe did not amplify.