Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 541021
Title Precision and accuracy of single-molecule FRET measurements—a multi-laboratory benchmark study
Author(s) Hellenkamp, Björn; Schmid, Sonja; Doroshenko, Olga; Opanasyuk, Oleg; Kühnemuth, Ralf; Rezaei Adariani, Soheila; Ambrose, Benjamin; Aznauryan, Mikayel; Barth, Anders; Birkedal, Victoria; Bowen, Mark E.; Chen, Hongtao; Cordes, Thorben; Eilert, Tobias; Fijen, Carel; Gebhardt, Christian; Götz, Markus; Gouridis, Giorgos; Gratton, Enrico; Ha, Taekjip; Hao, Pengyu; Hanke, Christian A.; Hartmann, Andreas; Hendrix, Jelle; Hildebrandt, Lasse L.; Hirschfeld, Verena; Hohlbein, Johannes; Hua, Boyang; Hübner, Christian G.; Kallis, Eleni; Kapanidis, Achillefs N.; Kim, Jae Yeol; Krainer, Georg; Lamb, Don C.; Lee, Nam Ki; Lemke, Edward A.; Levesque, Brié; Levitus, Marcia; McCann, James J.; Naredi-Rainer, Nikolaus; Nettels, Daniel; Ngo, Thuy; Qiu, Ruoyi; Robb, Nicole C.; Röcker, Carlheinz; Sanabria, Hugo; Schlierf, Michael; Schröder, Tim; Schuler, Benjamin; Seidel, Henning; Streit, Lisa; Thurn, Johann; Tinnefeld, Philip; Tyagi, Swati; Vandenberk, Niels; Vera, Andrés Manuel; Weninger, Keith R.; Wünsch, Bettina; Yanez-Orozco, Inna S.; Michaelis, Jens; Seidel, Claus A.M.; Craggs, Timothy D.; Hugel, Thorsten
Source Nature Methods : techniques for life scientists and chemists 15 (2018)9. - ISSN 1548-7091 - p. 669 - 676.
DOI https://doi.org/10.1038/s41592-018-0085-0
Department(s) Biophysics
EPS
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2018
Abstract

Single-molecule Förster resonance energy transfer (smFRET) is increasingly being used to determine distances, structures, and dynamics of biomolecules in vitro and in vivo. However, generalized protocols and FRET standards to ensure the reproducibility and accuracy of measurements of FRET efficiencies are currently lacking. Here we report the results of a comparative blind study in which 20 labs determined the FRET efficiencies (E) of several dye-labeled DNA duplexes. Using a unified, straightforward method, we obtained FRET efficiencies with s.d. between ±0.02 and ±0.05. We suggest experimental and computational procedures for converting FRET efficiencies into accurate distances, and discuss potential uncertainties in the experiment and the modeling. Our quantitative assessment of the reproducibility of intensity-based smFRET measurements and a unified correction procedure represents an important step toward the validation of distance networks, with the ultimate aim of achieving reliable structural models of biomolecular systems by smFRET-based hybrid methods.

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