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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 4.

Figure 4

Blockage of primer extension reactions catalyzed by pol η at the 4-OHEN-dC adduct site. Six products containing a single diastereoisomer of 4-OHEN-dC obtained as in Figure 3 were used for preparing their 38-mer oligomer templates as described in Materials and Methods. Using 150 fmol of the modified templates primed with 100 fmol of 32P-labeled 10-mer, primer extension reactions were carried out at 25 °C for 5 min in a buffer containing four dNTPs (100 μM each) and 10 fmol of pol η. One-third of the reaction mixture was subjected to denaturing PAGE (20% (w/v) polyacrylamide) in a 35 cm × 42 cm × 0.04 cm slab gel.