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. 2017 Oct 27;45(21):12057–12068. doi: 10.1093/nar/gkx990

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 2. — Preparation of the circular DNA template with 20 segment elements. (A) Schematical representation of the two-step ligation of several Outer Circle oligonucleotides (highlighted in green) into a single-stranded DNA circle. The addition of the Inner Circle oligonucleotides (ICs) results in the completion of the dsDNA nanostructure. (B) Native PAGE analysis of the ligation of the Outer Circle oligonucleotides. Lane 1: sequence OC01; lane 2: sequence OC02; lane 3: sequence OC03; lane 4: ligation of OC01 and OC02; lane 5: ligation of OC02 and OC03; lane 6: ligation of OC01 and OC03; lane 7: ligation of OC01, OC02 and OC03. (C) Native PAGE analysis of ligated and non-ligated Outer Circle oligonucleotides under different conditions for the binding of the Cap oligonucleotides. Annealing of the Cap sequences by heating-cooling treatment (lanes 1–4) or at room temperature (lanes 5–8). Resulting products were ligated for samples in lanes 1, 2, 5, 6. Caps were removed for lanes 1, 3, 5, 7.