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. 2018 Apr 18;8:6163. doi: 10.1038/s41598-018-24499-5

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© The Author(s) 2018

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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Biochemical analysis of supercoiled plasmids. (A) Plasmid DNA preparations were incubated with nuclease Bal-31 for increasing timespans, as indicated lanes 1–4, −SC plasmid; lanes 5–8, +SC plasmid. N mock-incubated −SC DNA; P mock-incubated highly +SC DNA, L linearized 3000 bp plasmid; M 1 kb DNA ladder. Over time, samples were removed, quenched by the addition of stop buffer and the products analysed on 1% agarose gel. (B) Incubation of plasmid DNA with glyoxal. Highly −SC and +Sc DNA preparations were incubated overnight in the absence or presence of increasing concentrations of glyoxal, as indicated, and analyzed on 0.8% agarose gel. The lanes 1–4 (−SC plasmid) and 1–8 (+SC plasmid) are cropped from different parts of the same gel. (C) Effect of temperature on glyoxal binding. Highly −SC and +SC DNA were incubated for 5 min. at indicated temperatures in the presence of 1 M glyoxal and analyzed by 0.8% agarose gel electrophoresis. The lanes 1–3 and 4–5 are cropped from different parts of the same gel.