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. 2018 Apr 25;8:6548. doi: 10.1038/s41598-018-24677-5

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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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Chemically modified ssDNA produced by aPCR. Accustart Taq polymerase HiFi is able to incorporate various chemically modified dNTPs into ssDNA produced with aPCR. (a) Triphosphate-modified nucleotide incorporation is realized by replacing canonical dNTPs with varying concentration of modified alpha-thiol dNTPs (4 bases). ssDNA production is effective for a replacement of dNTPs up to 75% (upper band: dsDNA 1,000 bp; lower band: ssDNA 1,000 nt). See Fig. S16 for higher percentage of dNTPs replacement. (b) dUTP replacement for dTTP up to 100% in asymmetric production of the ssDNA. Different parts of the same gel and exposure are shown and the split indicated by a white space. See Fig. S17 for the uncropped gel. (c) Cy5 fluorescently modified dCTP can be used to up to 10% replacement of canonical dCTP to be incorporated into the synthesized ssDNA (left). Merged image of ethidium bromide and Cy5 channels. See Fig. S19 for the full ethidium bromide- and Cy5-channel images. The efficiency of incorporation of Cy5-dCTP is quantified by fluorimetry and show approximately 2.5% modification of the Cy5 in ssDNA when 5% of Cy5-modified dCTP are used in aPCR for both 1,000 and 2,000 nt fragments (right). (d) Fluorescently modified ssDNA 2,000 nt is used to fold DNA nanoparticles. DNA stain and fluorescent agarose gel mobility shift assay showing folding of a fluorescent scaffolded DNA origami nanoparticle compared to non-fluorescent DNA nanoparticles.