Figure 8From: RNase H-dependent PCR (rhPCR): improved specificity and single nucleotide polymorphism detection using blocked cleavable primersApplication of rhPCR to determine genotype for 31 individuals at the SMAD7 rs4939827 SNP Locus. Human DNA samples from 31 individuals comprising 10 C/C homozygotes (circles), 10 T/T homozygotes (squares), and 11 C/T heterozygotes (triangles) were obtained from the Coriell Cell Repository and genotypes were determined using rhPCR. Reactions were run in triplicate using 2 ng of genomic DNA with SYBR® Green detection. Separate reactions using unmodified non-discriminatory control primers were performed to ensure that the amount of template was approximately equal in each case. A. Reactions were run in real-time mode for 45 cycles. Cq values for triplicate reactions were averaged and plotted according to the primer employed (rC vs. rU). B. Reactions were run in end-point mode for 35 cycles. The raw fluorescence measurements from each reaction are shown and plotted according to the primer employed (rC vs. rU).Back to article page