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. 2018 Apr 12;8:5925. doi: 10.1038/s41598-018-24417-9

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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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Plasmid capture in vitro via TFO-mediated triplex formation: 600 ng (≈140 fmol) of each plasmid (pAS03, pAS03.1, pAS03.2, pAS04) are mixed with 8 pmol of TFO-1 for 24 hr, followed by mixing with streptavidin magnetic beads. The plasmid fraction not bound to beads (unbound: UB) is recovered using a magnetic stand (lanes 1–6). After the washing steps, plasmid bound to the beads is released either by boiling (lanes 7–11) or by competitive elution using a biotin-containing buffer (lane 12). Following competitive elution, the beads are re-suspended in a buffer and further eluted by boiling (lane 13). All plasmids are supercoiled circular except for lanes 5 and 11 where the plasmid is linearized. The recovered samples are analysed by agarose gel electrophoresis. Similar results were obtained in three independent experiments and a representative gel picture is shown here. In addition, TFO-3 was shown to perform similarly (data not shown). The loading amounts are 15 ng (lanes 1–6), 30 ng (lanes 7–12), and 60 ng (lane 13) plasmid equivalent. Estimation of plasmid recovery compared to input is: pAS03, <1% (lane 7); pAS03.1, ≈50% (lane 8); pAS03.2, ≈60% (lane 9); pAS04 (lanes 10–12), ≈70%, as quantified by NIH ImageJ software. CC: closed circular. OC: open circular. Dimer: dimerized plasmid.