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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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2023-12-19

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Leitao_et_al_SI.pdf

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Postprint

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http://purl.org/coar/version/c_ab4af688f83e57aa

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embargo

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2023-12-19

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copyright

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Adobe PDF

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