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Electrical DNA Sequence Mapping Using Oligodeoxynucleotide Labels and Nanopores.

Accepted version
Peer-reviewed

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Type

Article

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Authors

Gularek, Felix 
Liu, Boyao 
Weinhold, Elmar 

Abstract

Identifying DNA species is crucial for diagnostics. For DNA identification, single-molecule DNA sequence mapping is an alternative to DNA sequencing toward fast point-of-care testing, which traditionally relies on targeting and labeling DNA sequences with fluorescent labels and readout using optical imaging methods. A nanopore is a promising sensor as a complement to optical mapping with advantages of electric measurement suitable for portable devices and potential for high resolution. Here, we demonstrate a high-resolution nanopore-based DNA sequence mapping by labeling specific short sequence motifs with oligodeoxynucleotides (ODNs) using DNA methyltransferase (MTase) and detecting them using nanopores. We successfully detected ODNs down to the size of 11 nucleotides without introducing extra reporters and resolved neighboring sites with a distance of 141 bp (∼48 nm) on a single DNA molecule. To accurately locate the sequence motif positions on DNA, a nanopore data analysis method is proposed by considering DNA velocity change through nanopores and using ensemble statistics to translate the time-dependent signals to the location information. Our platform enables high-resolution detection of small labels on DNA and high-accuracy localization of them for DNA species identification in an all-electrical format. The method presents an alternative to optical techniques relying on fluorescent labels and is promising for miniature-scale integration for diagnostic applications.

Description

Keywords

AdoMet analogue, DNA detection, DNA methyltransferase, nanopore sensing, single-molecule, Base Sequence, DNA, Nanopores, Nanotechnology, Oligodeoxyribonucleotides, Sequence Analysis, DNA

Journal Title

ACS Nano

Conference Name

Journal ISSN

1936-0851
1936-086X

Volume Title

15

Publisher

American Chemical Society (ACS)

Rights

All rights reserved
Sponsorship
European Research Council (647144)