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Magnetic torque tweezers: measuring torsional stiffness in DNA and RecA-DNA filaments

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Abstract

We introduce magnetic torque tweezers, which enable direct single-molecule measurements of torque. Our measurements of the effective torsional stiffness C of dsDNA indicated a substantial force dependence, with C = ∼40 nm at low forces up to C = ∼100 nm at high forces. The initial torsional stiffness of RecA filaments was nearly twofold larger than that for dsDNA, yet at moderate torques further build-up of torsional strain was prevented.

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Figure 1: Principle of magnetic torque tweezers and their operation.
Figure 2: Torque measurements for a 7.9 kbp dsDNA molecule in PBS buffer.
Figure 3: Measurements of RecA-DNA heteroduplex filaments.

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Acknowledgements

We thank M. Rojer for help with initial measurements, J. van der Does, D. de Roos and J. Beekman for help with instrumentation, and S. Hage and S. Donkers for providing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research (Nederlandse Organisatie voor Wetenschappelijk Onderzoek), Delft University of Technology and the European Science Foundation.

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Authors and Affiliations

  1. Department of Bionanoscience, Kavli Institute of Nanoscience, Faculty of Applied Sciences, Delft University of Technology, Delft, The Netherlands

    Jan Lipfert, Jacob W J Kerssemakers, Tessa Jager & Nynke H Dekker

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  1. Jan Lipfert
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  2. Jacob W J Kerssemakers
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  3. Tessa Jager
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  4. Nynke H Dekker
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Contributions

J.L., J.W.J.K. and N.H.D. designed the study, J.L. and T.J. performed the experiments, J.W.J.K. wrote the angular tracking routine and J.L. and N.H.D. wrote the manuscript.

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Correspondence to Nynke H Dekker.

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The authors declare no competing financial interests.

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Lipfert, J., Kerssemakers, J., Jager, T. et al. Magnetic torque tweezers: measuring torsional stiffness in DNA and RecA-DNA filaments. Nat Methods 7, 977–980 (2010). https://doi.org/10.1038/nmeth.1520

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