Additional File 6.
Figure S4. Mismatch discrimination using rhPCR with the mismatch positioned at the "-1" position relative to the RNA base. Sixteen synthetic oligonucleotide targets were employed where the base complementary to the single RNA residue in the blocked-cleavable primers was fixed (A, C, G, or T) and the base paired opposite position "-1" immediately 5'-to the RNA base in the primer was varied (A, C, G, or T). Likewise a set of 16 "rDDDDx" blocked-cleavable primers was employed where the RNA base was fixed (rA, rC, rG, or rU) and the base at the "-1" position was varied (A, C, G, or T). The target sequence and primers were otherwise the same as in Figure S3, except that the control non-discriminatory primer was one base shorter on the 3'-end. Assay conditions and calculations of ΔCq values were the same as in Figure 7 in the manuscript. All reactions were run in triplicate.
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Dobosy et al. BMC Biotechnology 2011 11:80 doi:10.1186/1472-6750-11-80