Print Email Facebook Twitter Invincible DNA tethers: Covalent DNA anchoring for enhanced temporal and force stability in magnetic tweezers experiments Title Invincible DNA tethers: Covalent DNA anchoring for enhanced temporal and force stability in magnetic tweezers experiments Author Janissen, R. Berghuis, B.A. Dulin, D. Wink, M. Van Laar, T. Dekker, N.H. Faculty Applied Sciences Department BN/Bionanoscience Date 2014-08-19 Abstract Magnetic tweezers are a powerful single-molecule technique that allows real-time quantitative investigation of biomolecular processes under applied force. High pulling forces exceeding tens of picoNewtons may be required, e.g. to probe the force range of proteins that actively transcribe or package the genome. Frequently, however, the application of such forces decreases the sample lifetime, hindering data acquisition. To provide experimentally viable sample lifetimes in the face of high pulling forces, we have designed a novel anchoring strategy for DNA in magnetic tweezers. Our approach, which exploits covalent functionalization based on heterobifunctional poly(ethylene glycol) crosslinkers, allows us to strongly tether DNA while simultaneously suppressing undesirable non-specific adhesion. A complete force and lifetime characterization of these covalently anchored DNA-tethers demonstrates that, compared to more commonly employed anchoring strategies, they withstand 3-fold higher pulling forces (up to 150 pN) and exhibit up to 200-fold higher lifetimes (exceeding 24 h at a constant force of 150 pN). This advance makes it possible to apply the full range of biologically relevant force scales to biomolecular processes, and its straightforward implementation should extend its reach to a multitude of applications in the field of single-molecule force spectroscopy. To reference this document use: http://resolver.tudelft.nl/uuid:ee24162e-211f-40e5-b997-aac5c08e0b38 DOI https://doi.org/10.1093/nar/gku677 Publisher Oxford University Press ISSN 0305-1048 Source http://nar.oxfordjournals.org/content/early/2014/08/19/nar.gku677.long Source Nucleic Acids Research, 42 (18), 2014 Part of collection Institutional Repository Document type journal article Rights (c) 2014 The Author(s)This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Files PDF Dekker_2014.pdf 1.28 MB Close viewer /islandora/object/uuid:ee24162e-211f-40e5-b997-aac5c08e0b38/datastream/OBJ/view