Research Data Leeds Repository

Citation

Chau, Chalmers C. and Radford, Sheena E. and Hewitt, Eric W. and Actis, Paolo (2020) Dataset for Macromolecular crowding enhances the detection of DNA and proteins by a solid-state nanopore. University of Leeds. [Dataset] https://doi.org/10.5518/841

Dataset description

Nanopore analysis of nucleic acid is now routine, but detection of proteins remains challenging. Here, we report the systematic characterization of the effect of macromolecular crowding on the detection sensitivity of a solid-state nanopore for circular and linearized DNA plasmids, globular proteins (beta-galactosidase), and filamentous proteins (alpha-synuclein amyloid fibrils). We observe a remarkable ca. 1000-fold increase in the molecule count for the globular protein beta-galactosidase and a 6-fold increase in peak amplitude for plasmid DNA under crowded conditions. We also demonstrate that macromolecular crowding facilitates the study of the topology of DNA plasmids and the characterization of amyloid fibril preparations with different length distributions. A remarkable feature of this method is its ease of use; it simply requires the addition of a macromolecular crowding agent to the electrolyte. We therefore envision that macromolecular crowding can be applied to many applications in the analysis of biomolecules by solid-state nanopores.

Keywords:	Nanopore, nanopipette, macromolecular crowding, single-molecule, DNA, protein, amyloid fibril.
Subjects:	H000 - Engineering > H600 - Electronic & electrical engineering
Divisions:	Faculty of Biological Sciences > Astbury Centre for Structural Molecular Biology Faculty of Biological Sciences > School of Molecular and Cellular Biology Faculty of Engineering and Physical Sciences > School of Electronic and Electrical Engineering
Related resources:	Location Type https://doi.org/10.1021/acs.nanolett.0c02246 Publication https://eprints.whiterose.ac.uk/162126/ Publication
License:	Creative Commons Attribution 4.0 International (CC BY 4.0)
Date deposited:	24 Jun 2020 14:29
URI:	https://archive.researchdata.leeds.ac.uk/id/eprint/699
Additional details Additional details Creators or authors: Creators ORCID Other ID Email Primary affiliation Primary affiliation ID Primary affiliation ID type Secondary affiliation Secondary affiliation ID Secondary affiliation ID type Chau, Chalmers C. https://orcid.org/0000-0002-3134-6798 University of Leeds https://ror.org/024mrxd33 ror Radford, Sheena E. https://orcid.org/0000-0002-3079-8039 University of Leeds https://ror.org/024mrxd33 ror Hewitt, Eric W. https://orcid.org/0000-0002-6238-6303 University of Leeds https://ror.org/024mrxd33 ror Actis, Paolo https://orcid.org/0000-0002-7146-1854 University of Leeds https://ror.org/024mrxd33 ror Type of data: Dataset Research funders: Wellcome Trust, ERC Grant numbers: 204963, 322408 Publication date: 2020 Resource language: English Metadata language: English Research project title: Macromolecular crowding enhances the detection of DNA and proteins by a solid-state nanopore Publisher: University of Leeds Contact email: p.actis@leeds.ac.uk Last Modified: 02 Sep 2024 12:10

Files

Documentation

Readme [759B] Readme [759B] Readme [759B] Security: Anyone File name: Readme.txt Content type: Documentation Mime-Type: text/plain License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 759B

Data

Amyloid Fibrils [92MB] Amyloid Fibrils [92MB] Amyloid Fibrils [92MB] Security: Anyone File name: Amyloid_Fibrils.zip Content type: Data Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 92MB

Βeta-galactosidase [297MB] Βeta-galactosidase [297MB] Βeta-galactosidase [297MB] Security: Anyone File name: Beta_galactosidase.zip Content type: Data Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 297MB

Circular DNA plasmid [254MB] Circular DNA plasmid [254MB] Circular DNA plasmid [254MB] Security: Anyone File name: Circular_DNA_plasmid.zip Content type: Data Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 254MB

Linearized DNA plasmid [48MB] Linearized DNA plasmid [48MB] Linearized DNA plasmid [48MB] Security: Anyone File name: Linearized_DNA_plasmid.zip Content type: Data Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 48MB

Supporting Figure 4, 5 [46MB] Supporting Figure 4, 5 [46MB] Supporting Figure 4, 5 [46MB] Security: Anyone File name: Supporting_Figure_4_5.zip Content type: Data Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 46MB