Volume 13, Issue 14 p. 1990-2001
Review

Advances in Quantitative FRET-Based Methods for Studying Nucleic Acids

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Søren Preus

Department of Chemistry, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen (Denmark)

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Assoc. Prof. L. Marcus Wilhelmsson

Corresponding Author

Assoc. Prof. L. Marcus Wilhelmsson

Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296 Göteborg (Sweden)

Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296 Göteborg (Sweden)Search for more papers by this author
First published: 31 August 2012
Citations: 124

Graphical Abstract

Accurate distance measurements: We provide a short overview of recent advances in using FRET as a quantitative ruler in nucleic acid systems, both in ensemble and in single-molecule studies. Topics include dye properties in DNA, alternative probes, accurate FRET measurements, quantitative data analysis, new FRET-based techniques and available software.

Abstract

Förster resonance energy transfer (FRET) is a powerful tool for monitoring molecular distances and interactions at the nanoscale level. The strong dependence of transfer efficiency on probe separation makes FRET perfectly suited for “on/off” experiments. To use FRET to obtain quantitative distances and three-dimensional structures, however, is more challenging. This review summarises recent studies and technological advances that have improved FRET as a quantitative molecular ruler in nucleic acid systems, both at the ensemble and at the single-molecule levels.