Figure 1.

Basic overview of the DPE-PCR assay. (A) DNA polymerase is incubated with a substrate consisting of pre-annealed Oligos 1 and 2. DNA polymerase extends only the 3′-end of Oligo 1 during a 20-min incubation at 37°C. Three microliters of the DPE reaction mixture is subsequently transferred into a hot start qPCR containing UDG. Prior to and during activation of Taq, UDG degrades the deoxyuridine within Oligo 2, leaving only a single-stranded product derived from DNA polymerase-mediated extension of Oligo 1. After activation of Taq, PCR-based amplification is initiated via reverse primer binding to the Oligo 1 extension product. (B) The sequence of a competitive internal control DNA is presented. The competitive internal control is present at 40 copies within each PCR.