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Single-Molecule Fluorescence of Nucleic Acids

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Fluorescence Spectroscopy and Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1076))

Abstract

Single-molecule fluorescence studies of nucleic acids are revolutionizing our understanding of fundamental cellular processes related to DNA and RNA processing mechanisms. Detailed molecular insights into DNA repair, replication, transcription, and RNA folding and function are continuously being uncovered by using the full repertoire of single-molecule fluorescence techniques. The fundamental reason behind the stunning growth in the application of single-molecule techniques to study nucleic acid structure and dynamics is the unmatched ability of single-molecule fluorescence, and mostly single-molecule FRET, to resolve heterogeneous static and dynamic populations and identify transient and low-populated states without the need for sample synchronization. New advances in DNA and RNA synthesis, post-synthetic dye-labeling methods, immobilization and passivation strategies, improved dye photophysics, and standardized analysis methods have enabled the implementation of single-molecule techniques beyond specialized laboratories. In this chapter, we introduce the practical aspects of applying single-molecule techniques to investigate nucleic acid structure, dynamics, and function.

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Author information

Authors and Affiliations

  1. SUPA School of Physics and Astronomy, University of St. Andrews, St. Andrews, Scotland, UK

    Kaley McCluskey & Euan Shaw

  2. Faculté des Sciences, RNA Group, Département de Biologie, Université de Sherbrooke, Sherbrooke, QC, Canada

    Daniel A. Lafontaine

  3. SUPA School of Physics and Astronomy, Biomedical Sciences Research Complex, University of St. Andrews, St. Andrews, Scotland, UK

    J. Carlos Penedo

Authors
  1. Kaley McCluskey
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  2. Euan Shaw
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  3. Daniel A. Lafontaine
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  4. J. Carlos Penedo
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Editor information

Editors and Affiliations

  1. KULeuven-Biomolecular Dynamics BioSCENTer, Leuven, Belgium

    Yves Engelborghs

  2. MicroSpectroscopy Centre, University of Wageningen Biochemistry, Wageningen, The Netherlands

    Antonie J.W.G. Visser

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McCluskey, K., Shaw, E., Lafontaine, D.A., Penedo, J.C. (2014). Single-Molecule Fluorescence of Nucleic Acids. In: Engelborghs, Y., Visser, A. (eds) Fluorescence Spectroscopy and Microscopy. Methods in Molecular Biology, vol 1076. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-649-8_35

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  • DOI: https://doi.org/10.1007/978-1-62703-649-8_35

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-648-1

  • Online ISBN: 978-1-62703-649-8

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