Figure 6
From: High accuracy genotyping directly from genomic DNA using a rolling circle amplification based assay
OCPs without hairpins, Factor V Leiden Mutation Ligation/ERCA Assay. Optimal assay conditions were as described in Methods. 1000 ng of MDA genomic DNA was incubated with probes for both alleles in the presence of ligase. ERCA mix was added and the reaction was followed in real time at 60°C in a Bio-Rad I-Cycler. The PCR baseline subtracted relative fluorescence units are shown as a function of time for both FAM (top row) and Cy3 (bottom row). Each trace (colored lines) corresponds to a different sample. Real time traces from reactions containing the same genotypes (either normal, heterozygous, or homozygous mutant) were grouped together according to genotype in each of the three columns. Genotypes were assigned as described in Methods.