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Nucleic acid purification from plants, animals and microbes in under 30 seconds

Nucleic acid amplification is a powerful molecular biology tool, although its use outside the modern laboratory environment is limited due to the relatively cumbersome methods required to extract nucleic acids from biological samples. To address this issue, we investigated a variety of materials for...

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Detalles Bibliográficos
Autores principales: Zou, Yiping, Mason, Michael Glenn, Wang, Yuling, Wee, Eugene, Turni, Conny, Blackall, Patrick J., Trau, Matt, Botella, Jose Ramon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5697807/
https://www.ncbi.nlm.nih.gov/pubmed/29161268
http://dx.doi.org/10.1371/journal.pbio.2003916
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author Zou, Yiping
Mason, Michael Glenn
Wang, Yuling
Wee, Eugene
Turni, Conny
Blackall, Patrick J.
Trau, Matt
Botella, Jose Ramon
author_facet Zou, Yiping
Mason, Michael Glenn
Wang, Yuling
Wee, Eugene
Turni, Conny
Blackall, Patrick J.
Trau, Matt
Botella, Jose Ramon
author_sort Zou, Yiping
collection PubMed
description Nucleic acid amplification is a powerful molecular biology tool, although its use outside the modern laboratory environment is limited due to the relatively cumbersome methods required to extract nucleic acids from biological samples. To address this issue, we investigated a variety of materials for their suitability for nucleic acid capture and purification. We report here that untreated cellulose-based paper can rapidly capture nucleic acids within seconds and retain them during a single washing step, while contaminants present in complex biological samples are quickly removed. Building on this knowledge, we have successfully created an equipment-free nucleic acid extraction dipstick methodology that can obtain amplification-ready DNA and RNA from plants, animals, and microbes from difficult biological samples such as blood and leaves from adult trees in less than 30 seconds. The simplicity and speed of this method as well as the low cost and availability of suitable materials (e.g., common paper towelling), means that nucleic acid extraction is now more accessible and affordable for researchers and the broader community. Furthermore, when combined with recent advancements in isothermal amplification and naked eye DNA visualization techniques, the dipstick extraction technology makes performing molecular diagnostic assays achievable in limited resource settings including university and high school classrooms, field-based environments, and developing countries.
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spelling pubmed-56978072017-11-30 Nucleic acid purification from plants, animals and microbes in under 30 seconds Zou, Yiping Mason, Michael Glenn Wang, Yuling Wee, Eugene Turni, Conny Blackall, Patrick J. Trau, Matt Botella, Jose Ramon PLoS Biol Methods and Resources Nucleic acid amplification is a powerful molecular biology tool, although its use outside the modern laboratory environment is limited due to the relatively cumbersome methods required to extract nucleic acids from biological samples. To address this issue, we investigated a variety of materials for their suitability for nucleic acid capture and purification. We report here that untreated cellulose-based paper can rapidly capture nucleic acids within seconds and retain them during a single washing step, while contaminants present in complex biological samples are quickly removed. Building on this knowledge, we have successfully created an equipment-free nucleic acid extraction dipstick methodology that can obtain amplification-ready DNA and RNA from plants, animals, and microbes from difficult biological samples such as blood and leaves from adult trees in less than 30 seconds. The simplicity and speed of this method as well as the low cost and availability of suitable materials (e.g., common paper towelling), means that nucleic acid extraction is now more accessible and affordable for researchers and the broader community. Furthermore, when combined with recent advancements in isothermal amplification and naked eye DNA visualization techniques, the dipstick extraction technology makes performing molecular diagnostic assays achievable in limited resource settings including university and high school classrooms, field-based environments, and developing countries. Public Library of Science 2017-11-21 /pmc/articles/PMC5697807/ /pubmed/29161268 http://dx.doi.org/10.1371/journal.pbio.2003916 Text en © 2017 Zou et al 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 use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Methods and Resources
Zou, Yiping
Mason, Michael Glenn
Wang, Yuling
Wee, Eugene
Turni, Conny
Blackall, Patrick J.
Trau, Matt
Botella, Jose Ramon
Nucleic acid purification from plants, animals and microbes in under 30 seconds
title Nucleic acid purification from plants, animals and microbes in under 30 seconds
title_full Nucleic acid purification from plants, animals and microbes in under 30 seconds
title_fullStr Nucleic acid purification from plants, animals and microbes in under 30 seconds
title_full_unstemmed Nucleic acid purification from plants, animals and microbes in under 30 seconds
title_short Nucleic acid purification from plants, animals and microbes in under 30 seconds
title_sort nucleic acid purification from plants, animals and microbes in under 30 seconds
topic Methods and Resources
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5697807/
https://www.ncbi.nlm.nih.gov/pubmed/29161268
http://dx.doi.org/10.1371/journal.pbio.2003916
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