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Assembly of Centromere Chromatin for Characterization by High-Speed Time-Lapse Atomic Force Microscopy

Part of the Methods in Molecular Biology book series (MIMB,volume 1814)

Abstract

Atomic force microscopy (AFM) is an imaging technique that enables single molecule characterization of biological systems at nanometer resolution. Imaging in ambient conditions can provide details of the conformational states and interactions of a population of molecules which is well complemented by single-molecule imaging of the systems dynamics using time-lapse AFM imaging, in which images are capture at rates of 10–15 frames per second in an aqueous buffer. Here we describe the assembly and preparation of nucleosomes containing centromere protein A (CENP-A) for AFM imaging in both static and time-lapse modes. The AFM imaging and data analysis techniques described enable characterization of the extent of DNA wrapping around the histone core and time-resolved visualization of the systems intrinsic dynamic behaviors.

Key words

  • High-speed atomic force microscopy
  • Chromatin dynamics
  • Nucleosomes
  • Centromere
  • CENP-A
  • Nanoimaging

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Acknowledgments

This work was supported by the National Science Foundation [grant MCB 1515346 to Y.L.L.].

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Correspondence to Micah P. Stumme-Diers or Yuri L. Lyubchenko .

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Stumme-Diers, M.P., Banerjee, S., Sun, Z., Lyubchenko, Y.L. (2018). Assembly of Centromere Chromatin for Characterization by High-Speed Time-Lapse Atomic Force Microscopy. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_14

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  • DOI: https://doi.org/10.1007/978-1-4939-8591-3_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8590-6

  • Online ISBN: 978-1-4939-8591-3

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