期刊
METHODS
卷 52, 期 2, 页码 192-200出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2010.06.010
关键词
Single molecule; Nucleic acid-protein complex; FRET; TIRE; Vesicle
资金
- National Science Foundation [0747285]
- National Institutes of Health [R01GM085116]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM085116] Funding Source: NIH RePORTER
- Div Of Molecular and Cellular Bioscience [0747285] Funding Source: National Science Foundation
Single-molecule fluorescence spectroscopy reveals the real time dynamics that occur during biomolecular interactions that would otherwise be hidden by the ensemble average. It also removes the requirement to synchronize reactions, thus providing a very intuitive approach to study kinetics of biological systems. Surface immobilization is commonly used to increase observation times to the minute time scale, but it can be detrimental if the sample interacts non-specifically with the surface. Here, we review detailed protocols to prevent such interactions by passivating the surface or by trapping the molecules inside surface immobilized lipid vesicles. Finally, we discuss recent examples where these methods were applied to study the dynamics of important cellular processes at the single-molecule level. (C) 2010 Elsevier Inc. All rights reserved.
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