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. Author manuscript; available in PMC: 2016 Mar 10.
Published in final edited form as: Biochemistry. 2015 Feb 26;54(9):1849–1857. doi: 10.1021/bi501491z

Figure 4.

Figure 4

An ICL forms between an abasic site and P located on opposing strands of the 35mer DNA duplex 2/3. A. The amount of 35mer-35mer ICL increases with time in a reaction that contains duplex 2/3 (lanes 3–7). No ICL forms in control reactions containing duplex 2/4 (lane 1) or single stranded 2 (lane 2). Reactions contained 0.25 µM DNA and were carried out at 37 °C and pH 6.5. Samples were reduced with NaBH4 prior to separation on a 20% denaturing polyacrylamide gel. The 17mer that is visible in all lanes arises from nonenzymatic β-elimination of 2 that occurs during preparation and storage of this oligonucleotide, and its amount (6%) remains constant during the reaction courses. B. The ratio of ICL to unlinked 2/3 at time points in A and duplicate reactions is shown. An end point ratio of 0.047 and kobs of 0.034 min−1 for the approach to the end point were obtained from an exponential fit to the data.