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research article

Spatially multiplexed single-molecule translocations through a nanopore at controlled speeds

Leitao, S. M.  
•
Navikas, V.  
•
Miljkovic, H.  
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June 19, 2023
Nature Nanotechnology

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. In single-molecule characterization, the near-infinite re-read capability on the same region of interest has the potential to unlock greater sensing capacity. A nanopore-based method, named scanning ion conductance spectroscopy, provides complete control over the translocation speed and nanopore position along a selected region and can detect a single 3 angstrom gap in a long strand of DNA.

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Type
research article
DOI
10.1038/s41565-023-01412-4
Web of Science ID

WOS:001010653400001

Author(s)
Leitao, S. M.  
Navikas, V.  
Miljkovic, H.  
Drake, B.  
Marion, S.  
Pistoletti Blanchet, G.  
Chen, K.
Mayer, S. F.  
Keyser, U. F.
Kuhn, A.
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Date Issued

2023-06-19

Published in
Nature Nanotechnology
Subjects

Nanoscience & Nanotechnology

•

Materials Science, Multidisciplinary

•

Science & Technology - Other Topics

•

Materials Science

•

trans location

•

dna

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microscopy

•

platform

•

velocity

•

noise

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBEN  
LBNI  
Available on Infoscience
July 17, 2023
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/199165
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