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. Author manuscript; available in PMC: 2017 Oct 31.
Published in final edited form as: Biochemistry. 2017 Jun 27;56(27):3415–3421. doi: 10.1021/acs.biochem.7b00386

Figure 2.

Figure 2

Cy3P/BioT DNA substrate for SSB. The duplex region is identical to that described previously for RPA1 and the size (29 bp) is in agreement with the requirements for assembly of a PCNA ring onto DNA by RFC2,3. The length (70 nt) of the 5′ polyT ssDNA overhang shown in red will accommodate only a single SSB molecule at 200 mM ionic strength and permits RFC-catalyzed loading of PCNA. When pre-bound to Neutravidin, the biotin attached to the 3′-end of the template strand prevents loaded PCNA from sliding off the duplex end. A Cy3 dye attached to the 5′end of the primer strand serves as the FRET donor.