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. 2019 Sep 25;9:13820. doi: 10.1038/s41598-019-49843-1

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© The Author(s) 2019

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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Site-specific attachment of DNA handles to a protein is necessary for force spectroscopy using a double-dumbbell optical tweezers set up. A small DNA oligo is covalently linked to the protein and hybridized to single-stranded overhangs in the DNA handles. The trapped beads can then be moved apart to exert a force on the protein.