Figure 10
From: High accuracy genotyping directly from genomic DNA using a rolling circle amplification based assay
Factor V Leiden Mutation: 87 different patient samples, 3 different genotypes. Ligation/ERCA genotyping results visualized as both a real-time assay and as an end-point. Assay conditions were as described in Methods for specific genotyping assay targets. 1000 ng of WGA genomic DNA was incubated with OCPs for both alleles in the presence of ligase. ERCA reactions were monitored in real time in the Bio-Rad I-cycler. Each colored trace in the real time charts corresponds to a different sample. At the end of the assay, approximately one-hour after addition of DNA polymerase, the intensity of both FAM and Cy3 fluorescence were measured for each sample in a fluorescent plate reader. End-point genotyping results were analyzed by the fuzzy cluster program and grouped automatically into genotypes as described in Methods.