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.
CITATION STYLE
Geffroy, L., Mangeol, P., Bizebard, T., & Bockelmann, U. (2018). RNA unzipping and force measurements with a dual optical trap. In Methods in Molecular Biology (Vol. 1665, pp. 25–41). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7271-5_2
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