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A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types

A method for the extraction of nucleic acids from a wide range of environmental samples was developed. This method consists of several modules, which can be individually modified to maximize yields in extractions of DNA and RNA or separations of DNA pools. Modules were designed based on elaborate te...

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Autores principales: Lever, Mark A., Torti, Andrea, Eickenbusch, Philip, Michaud, Alexander B., Šantl-Temkiv, Tina, Jørgensen, Bo Barker
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4436928/
https://www.ncbi.nlm.nih.gov/pubmed/26042110
http://dx.doi.org/10.3389/fmicb.2015.00476
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author Lever, Mark A.
Torti, Andrea
Eickenbusch, Philip
Michaud, Alexander B.
Šantl-Temkiv, Tina
Jørgensen, Bo Barker
author_facet Lever, Mark A.
Torti, Andrea
Eickenbusch, Philip
Michaud, Alexander B.
Šantl-Temkiv, Tina
Jørgensen, Bo Barker
author_sort Lever, Mark A.
collection PubMed
description A method for the extraction of nucleic acids from a wide range of environmental samples was developed. This method consists of several modules, which can be individually modified to maximize yields in extractions of DNA and RNA or separations of DNA pools. Modules were designed based on elaborate tests, in which permutations of all nucleic acid extraction steps were compared. The final modular protocol is suitable for extractions from igneous rock, air, water, and sediments. Sediments range from high-biomass, organic rich coastal samples to samples from the most oligotrophic region of the world's oceans and the deepest borehole ever studied by scientific ocean drilling. Extraction yields of DNA and RNA are higher than with widely used commercial kits, indicating an advantage to optimizing extraction procedures to match specific sample characteristics. The ability to separate soluble extracellular DNA pools without cell lysis from intracellular and particle-complexed DNA pools may enable new insights into the cycling and preservation of DNA in environmental samples in the future. A general protocol is outlined, along with recommendations for optimizing this general protocol for specific sample types and research goals.
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spelling pubmed-44369282015-06-03 A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types Lever, Mark A. Torti, Andrea Eickenbusch, Philip Michaud, Alexander B. Šantl-Temkiv, Tina Jørgensen, Bo Barker Front Microbiol Microbiology A method for the extraction of nucleic acids from a wide range of environmental samples was developed. This method consists of several modules, which can be individually modified to maximize yields in extractions of DNA and RNA or separations of DNA pools. Modules were designed based on elaborate tests, in which permutations of all nucleic acid extraction steps were compared. The final modular protocol is suitable for extractions from igneous rock, air, water, and sediments. Sediments range from high-biomass, organic rich coastal samples to samples from the most oligotrophic region of the world's oceans and the deepest borehole ever studied by scientific ocean drilling. Extraction yields of DNA and RNA are higher than with widely used commercial kits, indicating an advantage to optimizing extraction procedures to match specific sample characteristics. The ability to separate soluble extracellular DNA pools without cell lysis from intracellular and particle-complexed DNA pools may enable new insights into the cycling and preservation of DNA in environmental samples in the future. A general protocol is outlined, along with recommendations for optimizing this general protocol for specific sample types and research goals. Frontiers Media S.A. 2015-05-19 /pmc/articles/PMC4436928/ /pubmed/26042110 http://dx.doi.org/10.3389/fmicb.2015.00476 Text en Copyright © 2015 Lever, Torti, Eickenbusch, Michaud, Šantl-Temkiv and Jørgensen. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Lever, Mark A.
Torti, Andrea
Eickenbusch, Philip
Michaud, Alexander B.
Šantl-Temkiv, Tina
Jørgensen, Bo Barker
A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types
title A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types
title_full A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types
title_fullStr A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types
title_full_unstemmed A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types
title_short A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types
title_sort modular method for the extraction of dna and rna, and the separation of dna pools from diverse environmental sample types
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4436928/
https://www.ncbi.nlm.nih.gov/pubmed/26042110
http://dx.doi.org/10.3389/fmicb.2015.00476
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