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Spatially multiplexed single-molecule translocations through a nanopore at controlled speeds.
Leitao, S M; Navikas, V; Miljkovic, H; Drake, B; Marion, S; Pistoletti Blanchet, G; Chen, K; Mayer, S F; Keyser, U F; Kuhn, A; Fantner, G E; Radenovic, A.
Afiliación
  • Leitao SM; Laboratory for Bio- and Nano-Instrumentation, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland.
  • Navikas V; Laboratory of Nanoscale Biology, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland.
  • Miljkovic H; Laboratory of Nanoscale Biology, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland.
  • Drake B; Laboratory for Bio- and Nano-Instrumentation, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland.
  • Marion S; Laboratory of Nanoscale Biology, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland.
  • Pistoletti Blanchet G; Laboratory of Molecular Biology, Institute of Life Technologies, School of Engineering, HES-SO Valais-Wallis, Sion, Switzerland.
  • Chen K; Central Environmental Laboratory, Institute of Environmental Engineering, ENAC, EPFL, Sion, Switzerland.
  • Mayer SF; Cavendish Laboratory, University of Cambridge, Cambridge, UK.
  • Keyser UF; Laboratory of Nanoscale Biology, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland.
  • Kuhn A; Cavendish Laboratory, University of Cambridge, Cambridge, UK.
  • Fantner GE; Laboratory of Molecular Biology, Institute of Life Technologies, School of Engineering, HES-SO Valais-Wallis, Sion, Switzerland.
  • Radenovic A; Laboratory for Bio- and Nano-Instrumentation, Institute of Bioengineering, School of Engineering, EPFL, Lausanne, Switzerland. georg.fantner@epfl.ch.
Nat Nanotechnol ; 18(9): 1078-1084, 2023 09.
Article en En | MEDLINE | ID: mdl-37337057
In current nanopore-based label-free single-molecule sensing technologies, stochastic processes influence the selection of translocating molecule, translocation rate and translocation velocity. As a result, single-molecule translocations are challenging to control both spatially and temporally. Here we present a method using a glass nanopore mounted on a three-dimensional nanopositioner to spatially select molecules, deterministically tethered on a glass surface, for controlled translocations. By controlling the distance between the nanopore and glass surface, we can actively select the region of interest on the molecule and scan it a controlled number of times and at a controlled velocity. Decreasing the velocity and averaging thousands of consecutive readings of the same molecule increases the signal-to-noise ratio by two orders of magnitude compared with free translocations. We demonstrate the method's versatility by assessing DNA-protein complexes, DNA rulers and DNA gaps, achieving down to single-nucleotide gap detection.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Nanoporos Idioma: En Revista: Nat Nanotechnol Año: 2023 Tipo del documento: Article País de afiliación: Suiza Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Nanoporos Idioma: En Revista: Nat Nanotechnol Año: 2023 Tipo del documento: Article País de afiliación: Suiza Pais de publicación: Reino Unido