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Visualization of Recombinant DNA and Protein Complexes Using Atomic Force Microscopy
Published on: July 18, 2011
Visualization of DNA and protein-DNA complexes with atomic force microscopy
Yuri L Lyubchenko1, Alexander A Gall, Luda S Shlyakhtenko
1Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE, USA.
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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