FIGURE 2.

Miscoding specificities of rG, 8-oxo-rG, and 8-oxo-dG during translesion synthesis. A, overview of two-phase PAGE analysis. Unmodified or modified 38-mer template DNA was annealed to an Alexa 546-labeled 10-mer primer. Primer extension reactions catalyzed by 50 fmol of human pol κ (B) or pol η (C) were performed for 30 min at 25 °C in the presence of dNTPs. The fully extended products were extracted from the polyacrylamide gel. Then, the extracted products were annealed with a complementary 38-mer, digested with EcoRI, and loaded onto a two-phase polyacrylamide gel. To analyze miscoding properties, the mobility of the products were compared with those of Alexa 546-labeled 18-mer standard oligonucleotides containing dC (18C), dA (18A), dG (18G), or dT (18T) opposite the lesion and one- (Δ¹) or two-base (Δ²) deletions. The miscoding properties (%) were quantified and presented as the mean ± S.E. of at least two independent experiments under ”Results.“