We describe a method to extract quantitative information on DNA structural and configurational properties from high-resolution topographic maps recorded by atomic force microscopy (AFM). DNA molecules are deposited on mica surfaces from an aqueous solution, carefully dehydrated, and imaged in air in Tapping Mode. Upon extraction of the spatial coordinates of the DNA backbones from AFM images, several parameters characterizing DNA structure and configuration can be calculated. Here, we explain how to obtain the distribution of contour lengths, end-to-end distances, and gyration radii. This modular protocol can be also used to characterize other statistical parameters from AFM topographies.

Characterization of structural and configurational properties of DNA by atomic force microscopy / A. Meroni, F. Lazzaro, M. Muzi-Falconi, A. Podestà (METHODS IN MOLECULAR BIOLOGY). - In: Genome Instability : Methods and Protocols / [a cura di] M. Muzi-Falconi, G.W. Brown. - [s.l] : Humana, 2018. - ISBN 9781493973057. - pp. 557-573

Characterization of structural and configurational properties of DNA by atomic force microscopy

A. Meroni;F. Lazzaro;M. Muzi-Falconi;A. Podestà
2018

Abstract

We describe a method to extract quantitative information on DNA structural and configurational properties from high-resolution topographic maps recorded by atomic force microscopy (AFM). DNA molecules are deposited on mica surfaces from an aqueous solution, carefully dehydrated, and imaged in air in Tapping Mode. Upon extraction of the spatial coordinates of the DNA backbones from AFM images, several parameters characterizing DNA structure and configuration can be calculated. Here, we explain how to obtain the distribution of contour lengths, end-to-end distances, and gyration radii. This modular protocol can be also used to characterize other statistical parameters from AFM topographies.
Atomic force microscope/microscopy (AFM); DNA; DNA conformation; Mica; Molecular Biology; Genetics
Settore BIO/11 - Biologia Molecolare
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
Book Part (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/551592
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