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. 2018:1665:25-41.
doi: 10.1007/978-1-4939-7271-5_2.

RNA Unzipping and Force Measurements with a Dual Optical Trap

Laurent Geffroy  1 , Pierre Mangeol  1   2 , Thierry Bizebard  3 , Ulrich Bockelmann  4
Affiliations

RNA Unzipping and Force Measurements with a Dual Optical Trap

Laurent Geffroy et al. Methods Mol Biol. 2018.

Abstract

In order to mechanically unfold a single RNA molecule, an RNA/DNA hybrid construction is prepared which allows specific attachment to two micrometer-sized beads. A dual-beam optical trap thus holding the construct in solution captures the beads separately. Unfolding of a molecule is obtained by increasing the distance between the traps, one trap being slowly moved while the other is held fixed. Force is measured to sub-piconewton precision by back focal plane interferometry of the bead in the fixed trap. The experiment allows us to measure structure and base-sequence-dependent force signals. In this chapter, important technical aspects of this type of single-molecule force measurements are considered.

Keywords: DNA; Force; Molecular construction; Optical trap; RNA; Single-molecule; Unzipping.

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