Locked nucleic acid oligomers as handles for single molecule manipulation.
Bottom Line: A simple, minimally-perturbative labeling strategy would significantly broaden the possible applications of SMM experiments, perhaps even allowing the study of native biomolecular structures.Our results show that LNA provides a simple, stable means to functionalize dsDNA for manipulation.We provide design rules that will facilitate their use in future experiments.
Affiliation: Materials Department, University of California, Santa Barbara, Santa Barbara, CA 93106, USA.Show MeSH
Related in: MedlinePlus
Mentions: Overview of strategy to assay LNA/DNA binding strength and specificity in a magnetic tweezer: (A) Biotin-labeled LNA oligomers bind to surface-bound DNA, and mediate the tethering of a 1 μm streptavidin-coated magnetic bead. The tether is extended by a known force through application of a magnetic field, and its length is measured by optical bead tracking. (B) Force/length relations for individual tethers are fit to theory; one representative fit is plotted in black. The fits return estimates of each tether's persistence length, lp, and contour length, Lc; the latter is an estimate of the LNA/DNA binding position. Multiplexed measurements permit elastic measurements of many tethers. Histograms of the fit contour lengths report on the characteristics of the NAA/DNA interaction. The 17 force/extension curves shown here correspond to the histogram (gray bars) in Figure 2A.
Affiliation: Materials Department, University of California, Santa Barbara, Santa Barbara, CA 93106, USA.