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DNA barcodes for rapid, whole genome, single-molecule analyses

We report an approach for visualizing DNA sequence and using these ‘DNA barcodes’ to search complex mixtures of genomic material for DNA molecules of interest. We demonstrate three applications of this methodology; identifying specific molecules of interest from a dataset containing gigabasepairs of...

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Detalles Bibliográficos
Autores principales: Wand, Nathaniel O, Smith, Darren A, Wilkinson, Andrew A, Rushton, Ashleigh E, Busby, Stephen J W, Styles, Iain B, Neely, Robert K
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6614835/
https://www.ncbi.nlm.nih.gov/pubmed/30918971
http://dx.doi.org/10.1093/nar/gkz212
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author Wand, Nathaniel O
Smith, Darren A
Wilkinson, Andrew A
Rushton, Ashleigh E
Busby, Stephen J W
Styles, Iain B
Neely, Robert K
author_facet Wand, Nathaniel O
Smith, Darren A
Wilkinson, Andrew A
Rushton, Ashleigh E
Busby, Stephen J W
Styles, Iain B
Neely, Robert K
author_sort Wand, Nathaniel O
collection PubMed
description We report an approach for visualizing DNA sequence and using these ‘DNA barcodes’ to search complex mixtures of genomic material for DNA molecules of interest. We demonstrate three applications of this methodology; identifying specific molecules of interest from a dataset containing gigabasepairs of genome; identification of a bacterium from such a dataset and, finally, by locating infecting virus molecules in a background of human genomic material. As a result of the dense fluorescent labelling of the DNA, individual barcodes of the order 40 kb pairs in length can be reliably identified. This means DNA can be prepared for imaging using standard handling and purification techniques. The recorded dataset provides stable physical and electronic records of the total genomic content of a sample that can be readily searched for a molecule or region of interest.
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spelling pubmed-66148352019-07-12 DNA barcodes for rapid, whole genome, single-molecule analyses Wand, Nathaniel O Smith, Darren A Wilkinson, Andrew A Rushton, Ashleigh E Busby, Stephen J W Styles, Iain B Neely, Robert K Nucleic Acids Res Methods Online We report an approach for visualizing DNA sequence and using these ‘DNA barcodes’ to search complex mixtures of genomic material for DNA molecules of interest. We demonstrate three applications of this methodology; identifying specific molecules of interest from a dataset containing gigabasepairs of genome; identification of a bacterium from such a dataset and, finally, by locating infecting virus molecules in a background of human genomic material. As a result of the dense fluorescent labelling of the DNA, individual barcodes of the order 40 kb pairs in length can be reliably identified. This means DNA can be prepared for imaging using standard handling and purification techniques. The recorded dataset provides stable physical and electronic records of the total genomic content of a sample that can be readily searched for a molecule or region of interest. Oxford University Press 2019-07-09 2019-03-28 /pmc/articles/PMC6614835/ /pubmed/30918971 http://dx.doi.org/10.1093/nar/gkz212 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Wand, Nathaniel O
Smith, Darren A
Wilkinson, Andrew A
Rushton, Ashleigh E
Busby, Stephen J W
Styles, Iain B
Neely, Robert K
DNA barcodes for rapid, whole genome, single-molecule analyses
title DNA barcodes for rapid, whole genome, single-molecule analyses
title_full DNA barcodes for rapid, whole genome, single-molecule analyses
title_fullStr DNA barcodes for rapid, whole genome, single-molecule analyses
title_full_unstemmed DNA barcodes for rapid, whole genome, single-molecule analyses
title_short DNA barcodes for rapid, whole genome, single-molecule analyses
title_sort dna barcodes for rapid, whole genome, single-molecule analyses
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6614835/
https://www.ncbi.nlm.nih.gov/pubmed/30918971
http://dx.doi.org/10.1093/nar/gkz212
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