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. Author manuscript; available in PMC: 2008 Aug 31.
Published in final edited form as: J Mol Biol. 2007 Jun 9;371(5):1151–1162. doi: 10.1016/j.jmb.2007.06.009

Figure 9.

Figure 9

Miscoding specificities of 4-OHEN-dC lesion in reactions catalyzed by the combination of pol η+ι or pol κ+ι. Using 150 fmol of a 4-OHEN-dC-modified and unmodified 38-mer template primed with 100 fmol of a 32P-labeled 12-mer primer, we conducted primer extension reactions at 25 °C for 30 min in a buffer containing four dNTPs (100 μM each) and either (a) pol η+ι (0–400 fmol for pol ι, 200 fmol for pol η) or (b) pol κΔC+ι (0–400 fmol for pol ι, 200 fmol for pol κΔC), as described in Materials and Methods. The extended reaction products (>26 bases long) produced on the unmodified and 4-OHEN-dC-modified templates were extracted following PAGE. The recovered oligodeoxynucleotides were annealed to an unmodified 38-mer and cleaved with EcoRI restriction enzyme, as described in Materials and Methods. Half of the reaction product from the unmodified template and the entire product from the 4-OHEN-dC-modified template were subjected to two-phased electrophoresis in a 20% (w/v) polyacrylamide 15 cm × 72 cm × 0.04 cm slab gel.