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. 2019 Jun 20;9:8853. doi: 10.1038/s41598-019-45356-z

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© The Author(s) 2019

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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Validation of the assay for alternative enzymes that digest ssDNA and generate dsDNA. (a) Mammalian Trex2 digestion of single stranded DNA quantified using the PG assay. Single-stranded 60-mer (orange), 40-mer (green) and 20-mer (purple) DNA substrates were treated with 100 and 200 nM Trex2. Robust digestion was observed in each case. (b) Time (seconds) until reaction completion based on the point at which the graphs plateau in (a). (c) Rate of digestion per nM Trex2. Trex2 degrades DNA at a similar rate irrespective of the length of the DNA substrate. (d) The polymerase activity of the Klenow fragment polymerase determined using the PG assay. Three substrates with a 20-, 40- and 60- and overhang with a total 80 bases were incubated with 1 nM Klenow fragment. (e) Calculated polymerisation rate of the Klenow fragment. A clear preference is shown for longer overhangs. Error bars represent SEM; n = 3 in all cases; ****p < 0.0001.