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. Author manuscript; available in PMC: 2019 Feb 1.
Published in final edited form as: DNA Repair (Amst). 2018 Jan 9;62:18–27. doi: 10.1016/j.dnarep.2018.01.004

Figure 1. Quantification of damaged plasmid via strand-specific primer extension/qPCR (SSPE-qPCR).

Figure 1

Repair of damaged plasmids is quantified using SSPE-qPCR: (1) Plasmid DNA is denatured, (2) a primer extension reaction is performed using primer R, (3) the denaturation/primer extension step is repeated and additional 7 cycles. After 8 cycles, primer L is added and Ct values determined using qPCR. The presence of an abasic site, cholesterol, or DPC (A) will cause Taq polymerase to stall, resulting in no full-length product strands. In contrast, when repair has occurred (B), Taq polymerase will produce full-length product strands containing the binding site for primer L.