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Single-Molecule FRET X

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Single Molecule Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2694))

Abstract

Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon that has been repurposed as a biophysical tool to measure nanometer distances. With FRET by DNA eXchange, or FRET X, many points of interest (POIs) in a single object can be probed, overcoming a major limitation of conventional single-molecule FRET. In FRET X, short fluorescently labeled DNA imager strands specifically and transiently bind their complementary docking strands on a target molecule, such that at most a single FRET pair is formed at each point in time and multiple POIs on a single molecule can be readily probed. Here, we describe the sample preparation, image acquisition, and data analysis for structural analysis of DNA nanostructures with FRET X.

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Declaration of Interests

C.J. and M.F. hold a patent on single-molecule FRET for protein characterization (patent number: WO2021049940).

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Authors and Affiliations

  1. Department of BioNanoScience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, The Netherlands

    Mike Filius, Raman van Wee & Chirlmin Joo

Authors
  1. Mike Filius
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  2. Raman van Wee
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  3. Chirlmin Joo
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Corresponding author

Correspondence to Chirlmin Joo .

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Editors and Affiliations

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

    Iddo Heller

  2. LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands

    David Dulin

  3. LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands

    Erwin J.G. Peterman

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Filius, M., van Wee, R., Joo, C. (2024). Single-Molecule FRET X. In: Heller, I., Dulin, D., Peterman, E.J. (eds) Single Molecule Analysis . Methods in Molecular Biology, vol 2694. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3377-9_10

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  • DOI: https://doi.org/10.1007/978-1-0716-3377-9_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3376-2

  • Online ISBN: 978-1-0716-3377-9

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