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Combined Magnetic Tweezers and Micro-mirror Total Internal Reflection Fluorescence Microscope for Single-Molecule Manipulation and Visualization

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Single Molecule Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1665))

Abstract

Magnetic tweezers is a versatile yet simple single-molecule manipulation technique that has been used to study a broad range of nucleic acids and nucleic acid-based molecular motors. In this chapter, we combine micro-mirror-based total internal reflection microscopy with a magnetic tweezers instrument, permitting simultaneous single-molecule visualization and mechanical manipulation. We provide a simple method to calibrate the evanescent wave penetration depth via supercoiling of DNA with a fluorescent nanodiamond-labeled magnetic bead and a complementary method employing a surface-immobilized fluorescent nanodiamond.

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Acknowledgements

We thank Ganesh Shenoy, Haksung Jung, and Rolf Swenson for providing biotinylated fluorescent nanodiamonds. We thank Gary Melvin for machining. This research was supported by the Intramural Research Program of the National Heart Lung and Blood Institute, National Institutes of Health.

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Authors and Affiliations

  1. Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 50, Room 3517, 50 South Drive, Bethesda, 20892, MD, USA

    Yeonee Seol & Keir C. Neuman

Authors
  1. Yeonee Seol
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  2. Keir C. Neuman
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Correspondence to Keir C. Neuman .

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Editors and Affiliations

  1. Department of Physics & Astronomy, Vrije Universiteit, Amsterdam, The Netherlands

    Erwin J. G. Peterman

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Seol, Y., Neuman, K.C. (2018). Combined Magnetic Tweezers and Micro-mirror Total Internal Reflection Fluorescence Microscope for Single-Molecule Manipulation and Visualization. In: Peterman, E. (eds) Single Molecule Analysis. Methods in Molecular Biology, vol 1665. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7271-5_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7271-5_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7270-8

  • Online ISBN: 978-1-4939-7271-5

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