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Methods Mol Biol. 2011;783:265-93. doi: 10.1007/978-1-61779-282-3_15.

Magnetic tweezers for single-molecule manipulation.

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  • 1Laboratory of Molecular Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.

Abstract

Magnetic tweezers provide a versatile tool enabling the application of force and torque on individual biomolecules. Magnetic tweezers are uniquely suited to the study of DNA topology and protein-DNA interactions that modify DNA topology. Perhaps due to its presumed simplicity, magnetic tweezers instrumentation has been described in less detail than comparable techniques. Here, we provide a comprehensive description and guide for the design and implementation of a magnetic tweezers instrument for single-molecule measurements of DNA topology and mechanics. We elucidate magnetic trap design, as well as microscope and illumination setup, and provide a simple LabVIEW-based real-time position tracking algorithm. In addition, we provide procedures for production of supercoilable DNA tethers, flow-cell design, and construction tips.

[PubMed - indexed for MEDLINE]
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