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Visualization of DNA and Protein–DNA Complexes with Atomic Force Microscopy

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Book cover Electron Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1117))

Abstract

This article describes sample preparation techniques for AFM imaging of DNA and protein–DNA complexes. The approach is based on chemical functionalization of the mica surface with aminopropyl silatrane (APS) to yield an APS-mica surface. This surface binds nucleic acids and nucleoprotein complexes in a wide range of ionic strengths, in the absence of divalent cations, and in a broad range of pH. The chapter describes the methodologies for the preparation of APS-mica surfaces and the preparation of samples for AFM imaging. The protocol for synthesis and purification of APS is also provided. The AFM applications are illustrated with examples of images of DNA and protein–DNA complexes.

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Acknowledgements

The authors thank the members of the Lyubchenko lab for their contribution to different parts of the paper. The work is supported by grants to Y.L.L. from the DOE (DE-FG02-08ER64579), NIH (P01 GM091743, 5 R01 GM096039-02), NSF (EPS—1004094), and the Nebraska Research Initiative (NRI).

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE, USA

    Yuri L. Lyubchenko & Luda S. Shlyakhtenko

  2. Cepheid, Bothell, WA, USA

    Alexander A. Gall

Authors
  1. Yuri L. Lyubchenko
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  2. Alexander A. Gall
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  3. Luda S. Shlyakhtenko
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Editor information

Editors and Affiliations

  1. University of Western Australia, Crawley, Australia

    John Kuo

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Lyubchenko, Y.L., Gall, A.A., Shlyakhtenko, L.S. (2014). Visualization of DNA and Protein–DNA Complexes with Atomic Force Microscopy. In: Kuo, J. (eds) Electron Microscopy. Methods in Molecular Biology, vol 1117. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-776-1_17

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  • DOI: https://doi.org/10.1007/978-1-62703-776-1_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-775-4

  • Online ISBN: 978-1-62703-776-1

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