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Single Molecule Imaging of DNA–Protein Interactions Using DNA Curtains

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DNA-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2599))

Abstract

Direct observation of enzymes that work to promote nucleic acid metabolism is a powerful approach to understanding their biochemical and biological properties. Over several years, fluorescent optical microscopy has developed as a powerful tool for watching biological pathways as they occur in real time. Here we describe DNA curtains as an optical microscopy tool that combines engineering, biochemistry, and single molecule imaging to make direct observations of enzymes as they work on DNA in real time. We will provide a detailed methodology of this approach including information about the setup of a basic TIRF microscope, assembly of flow chambers for imaging, and the protocol for making DNA curtains. Our goal is to help the reader better understand the technical approaches to DNA curtains and to better understand the biochemical and biological applications of this approach.

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Acknowledgments

The author would like to thank the members of the Crickard lab for critical reading of the manuscript. This research was funded by NIH Grant R35GM142457 (J.B.C).

Conflict of Interest

The author declares no competing conflict of interest.

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Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA

    J. Brooks Crickard

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  1. J. Brooks Crickard
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Correspondence to J. Brooks Crickard .

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Editors and Affiliations

  1. Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA

    Marcos Simoes-Costa

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Crickard, J.B. (2023). Single Molecule Imaging of DNA–Protein Interactions Using DNA Curtains. In: Simoes-Costa, M. (eds) DNA-Protein Interactions. Methods in Molecular Biology, vol 2599. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2847-8_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2847-8_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2846-1

  • Online ISBN: 978-1-0716-2847-8

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