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Methods. 2019 Jul 25. pii: S1046-2023(18)30306-2. doi: 10.1016/j.ymeth.2019.07.020. [Epub ahead of print]

Parallelized DNA tethered bead measurements to scrutinize DNA mechanical structure.

Author information

1
Laboratoire de Physique de l'Ecole Normale Supérieure, ENS, Université PSL, CNRS, Sorbonne Université, Université Paris-Diderot, Sorbonne Paris Cité, Paris, France.
2
Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
3
Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France. Electronic address: laurence.salome@ipbs.fr.

Abstract

Tethering beads to DNA offers a panel of single molecule techniques for the refined analysis of the conformational dynamics of DNA and the elucidation of the mechanisms of enzyme activity. Recent developments include the massive parallelization of these techniques achieved by the fabrication of dedicated nanoarrays by soft nanolithography. We focus here on two of these techniques: the Tethered Particle motion and Magnetic Tweezers allowing analysis of the behavior of individual DNA molecules in the absence of force and under the application of a force and/or a torque, respectively. We introduce the experimental protocols for the parallelization and discuss the benefits already gained, and to come, for these single molecule investigations.

KEYWORDS:

DNA conformational changes; Dynamic activity of DNA enzymes; High-throughput single-molecule techniques; Magnetic tweezers; Microcontact printing; Tethered particle motion

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