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. 2017 Jul 6;7:4756. doi: 10.1038/s41598-017-02578-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Processivity of Dbh and Sdbh. The ability of Dbh and Sdbh to extend a FAM-labeled primer (FAM-36 primer) annealed to single-stranded M13mp18 DNA from a single DNA binding event in the presence of a DNA trap. In lanes 1–4, the pol was preincubated with the primer-template DNA (12.5 nM) and the reactions were initiated by the addition of dNTPs (200 μM) and excess sperm DNA trap. As a control (lanes 5–8), the pol was preincubated with the primer-template DNA and the herring sperm DNA trap, and reactions were initiated by the addition of dNTPs. The intensity along the length of the lane was quantified and then the marker standards were used to assign a basepair length through the lanes using ImageQuant software. From there the greatest abundant product could be measured as the processivity. The unlabeled marker standards were visualized by SYBR stain independently and showed the length (nt) on the left of the first lane. (Note: the synthesized primer was 36 nt, but the FAM label altered its electrophoretic mobility, such that it ran at about 80 nt). See Supplemental S1 for full-length image.