4.3 Article

Simple horizontal magnetic tweezers for micromanipulation of single DNA molecules and DNA-protein complexes

Journal

BIOTECHNIQUES
Volume 60, Issue 1, Pages 21-27

Publisher

FUTURE SCI LTD
DOI: 10.2144/000114369

Keywords

single molecule-micromanipulation; magnetic tweezers; differential measurement; magnetic trap

Funding

  1. NIH [R01 AA17626]
  2. Vitreous State Laboratory
  3. Catholic University of America

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We report the development of a simple-to-implement magnetic force transducer that can apply a wide range of piconewton (pN) scale forces on single DNA molecules and DNA-protein complexes in the horizontal plane. The resulting low-noise force-extension data enable very high-resolution detection of changes in the DNA tether's extension: similar to 0.05 pN in force and < 10 nm change in extension. We have also verified that we can manipulate DNA in near equilibrium conditions through the wide range of forces by ramping the force from low to high and back again, and observing minimal hysteresis in the molecule's force response. Using a calibration technique based on Stokes' drag law, we have confirmed our force measurements from DNA force-extension experiments obtained using the fluctuation-dissipation theorem applied to transverse fluctuations of the magnetic microsphere. We present data on the force-distance characteristics of a DNA molecule complexed with histones. The results illustrate how the tweezers can be used to study DNA binding proteins at the single molecule level.

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