Visualization of DNA and protein-DNA complexes with atomic force microscopy

Yuri L. Lyubchenko, Alexander A. Gall, Luda S. Shlyakhtenko

Research output: Chapter in Book/Report/Conference proceedingChapter

19 Scopus citations

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 purifi cation of APS is also provided. The AFM applications are illustrated with examples of images of DNA and protein-DNA complexes.

Original languageEnglish (US)
Title of host publicationElectron Microscopy
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages367-384
Number of pages18
ISBN (Print)9781627037754
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1117
ISSN (Print)1064-3745

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Keywords

  • AFM
  • Atomic force microscopy
  • DNA structure and dynamics
  • Mica functionalization
  • Protein-DNA complexes
  • Silanes
  • Silatranes
  • Surface chemistry

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Lyubchenko, Y. L., Gall, A. A., & Shlyakhtenko, L. S. (2014). Visualization of DNA and protein-DNA complexes with atomic force microscopy. In Electron Microscopy: Methods and Protocols (pp. 367-384). (Methods in Molecular Biology; Vol. 1117). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-776-1_17