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. 2023 Feb 6;14:631. doi: 10.1038/s41467-023-36373-8

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© The Author(s) 2023

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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Fig. 1 — a Model of a DNA duplex⁶⁶ with enlarged frame showing stacked adjacent bases. b Design of two duplexes differing by a single-base stacking interaction. c Free-energy diagram of a DNA duplex with and without a terminal base stack. The base stack primarily increases the activation energy from E_A1 to E_A2, with the difference representing the free energy of the single terminal base stack, ΔG_stack. d External force lowers the activation barriers and prevents rebinding, causing exponential dissociation that can be experimentally measured and used to calculate stacking free energy.