title: Single molecule FRET data analysis procedures for FRET efficiency determination: probing the conformations of nucleic acid structures.
abstract: Single molecule FRET microscopy is an attractive technique for studying structural dynamics and conformational diversity of nucleic acid structures. Some of its strengths are that it can follow structural changes on a fast time scale and identify conformation distributions arising from dynamic or static population heterogeneity. Here, we give a description of the experiment and data analysis procedures of this method and detail what parameters are needed for FRET efficiency calculation. Using single molecule FRET data obtained on G-quadruplex DNA structures that exhibit large conformation diversity, we illustrate that the shape of the FRET distribution changes depending on what parameters are included in the data analysis procedure.
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark.
authors.forename: Asger Christian
authors.initials: AC
authors.lastname: Krüger
authors.forename: Victoria
authors.initials: V
authors.lastname: Birkedal
mesh_terms: D004247:DNA / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D054856:G-Quadruplexes
chemical_list: D004247:DNA
keywords: Fluorescence resonance energy transfer; G-quadruplex DNA structures; Nucleic acid structures; Single molecule microscopy