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Rapid assessment of site specific DNA methylation through resistive pulse sensing

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posted on 04.04.2018 by Matthew Healey, William Rowe, Sofia Siati, Muttuswamy Sivakumaran, Mark Platt
Many diseases are defined by patterns of DNA methylation which result in aberrant gene expression. We present a rapid assay based upon resistive pulse sensing, RPS, to characterize sequence specific DNA methylation sites in genomic DNA. We modify the surface of superparamagnetic beads, SPBs, with DNA (capture probe). The particles are added to solution where they bind to and extract sequence specific DNA (target DNA). The target loaded SPBs are then incubated with antibodies which bind to the methylation sites, and the velocity of the SPBs through the nanopore reveals the number and location of the epigenetic markers within the target. The approach is capable of distinguishing between different methylation sites within a DNA promoter region. Crucially the approach is not dependent on accurate sequencing of assayed DNA, with genomic regions targeted through complementary probes. As such the number of stages and reagents costs are low and the assay is complete in under 60 min which includes the incubation and run times. The format also allows simultaneous quantification of number of copies of methylated DNA, and we illustrate this with a dose response curve.

History

School

  • Science

Department

  • Chemistry

Published in

ACS Sensors

Citation

HEALEY, M.J. ... et al, 2018. Rapid assessment of site specific DNA methylation through resistive pulse sensing. ACS Sensors, 3 (3), pp.655-660.

Publisher

© American Chemical Society

Version

AM (Accepted Manuscript)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/

Acceptance date

14/03/2018

Publication date

2018-03-07

Notes

This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Sensors, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://pubs.acs.org/doi/10.1021/acssensors.7b00935.

ISSN

2379-3694

Language

en

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