Cargando…
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...
Autores principales: | , , , , , , , |
---|---|
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 |
_version_ | 1783280673357824000 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5697807 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT zouyiping nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT masonmichaelglenn nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT wangyuling nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT weeeugene nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT turniconny nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT blackallpatrickj nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT traumatt nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds AT botellajoseramon nucleicacidpurificationfromplantsanimalsandmicrobesinunder30seconds |