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Methods Mol Biol. 2015;1259:257-73. doi: 10.1007/978-1-4939-2214-7_16.

Constructing a magnetic tweezers to monitor RNA translocation at the single-molecule level.

Author information

1
Centre de Biochimie Structurale, CNRS UMR5048, 29 rue de Navacelles, 34090, Montpellier, France.

Abstract

Single-molecule methods have become an invaluable tool in the investigation of the mechanisms of nucleic-acid motors. Magnetic tweezers is a single-molecule manipulation technique that permits the real-time measurement of enzyme activities on single nucleic-acid molecules at high-resolution, high-throughput, and inherently constant force. Here, we describe several aspects of the implementation of magnetic tweezers, with special emphasis on the construction of a simple magnetic trap and, in particular, on the detailed description of image analysis methods to measure the extension changes in nucleic-acid molecules induced by protein activity. Finally, we carefully describe the steps involved in performing a full magnetic tweezers experiment.

PMID:
25579591
DOI:
10.1007/978-1-4939-2214-7_16
[Indexed for MEDLINE]

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