Journal
METHODS
Volume 49, Issue 2, Pages 112-117Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2009.04.017
Keywords
Single-molecule; FRET; TIRF; RNA; Kinetics; Folding
Funding
- National Science Foundation [0747285]
- National Institutes of Health [R01GM085116]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM085116] Funding Source: NIH RePORTER
- Direct For Biological Sciences [0747285] Funding Source: National Science Foundation
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Single-molecule fluorescence resonance energy transfer (smFRET) microscopy has become an increasingly popular tool to study the structural dynamics of RNA molecules. It reveals, in real time, the structural dynamics of these molecules that would be otherwise hidden in ensemble-averaged measurements. Here we present a detailed protocol for performing smFRET using total internal reflection fluorescence microscopy, including RNA preparation, optical setup, separation of the dual color channels, sample immobilization and data acquisition and analysis. (c) 2009 Published by Elsevier Inc.
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