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. Author manuscript; available in PMC: 2015 Aug 15.
Published in final edited form as: Bioorg Med Chem. 2014 Jun 2;22(16):4399–4406. doi: 10.1016/j.bmc.2014.05.049

Figure 7. Fluorescently-labeled oligonucleotides for kinetic analysis of DNA unwinding and translocation on ssDNA.

Figure 7

A. In the example shown, a fluorescent probe is placed onto the end of a DNA substrate and the complementary strand contains a fluorescence quencher. Upon helicase-catalyzed separation of the strands, an increase in fluorescence can be observed. B. Fluorescently-labeled oligonucleotides serve as a substrate for measuring translocation on ssDNA. When a helicase translocates to the end of the DNA, a change in fluorescence can be observed due to the interaction between the helicase and the fluorescent probe. The time required to reach the end of different length oligonucleotides can be used to determine the rate of translocation.