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. 2014 Aug 26;42(16):10655–10667. doi: 10.1093/nar/gku774

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — (A) DNA binding by clamp loaders. Duplex DNA spirals inside the cap of the clamp loader, with the single-stranded portion exiting between a gap in the clamp loader subunits (bacteriophage T4 clamp loader, PDB ID: 3U60 (15)). (B) A diagram showing the sequential mixing scheme in the stopped-flow. A solution of clamp loader in syringe 1, is mixed with fluorescent clamp and ATP from syringe 2 for 4 s to form an ATP·clamp loader·clamp complex. This complex is mixed with DNA, excess unlabeled clamp, and ATP. When SSB or RPA are present in reactions, they are included in syringe 3 with DNA to form SSB·DNA complexes. (C) Diagrams of the DNA structures used in these studies. All DNA structures are composed of two 60-mer oligonucleotides, annealed to produce a 30-nt duplex region, with two symmetrical 30-nt overhangs on either end of the duplex region. RPA was substituted for SSB in experiments with RFC and PCNA.