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A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments

Recovering high quality genomic DNA from environmental samples is a crucial primary step to understand the genetic, metabolic, and evolutionary characteristics of microbial communities through molecular ecological approaches. However, it is often challenging because of the difficulty of effective ce...

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Autores principales: Natarajan, Vengadesh Perumal, Zhang, Xinxu, Morono, Yuki, Inagaki, Fumio, Wang, Fengping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4917542/
https://www.ncbi.nlm.nih.gov/pubmed/27446026
http://dx.doi.org/10.3389/fmicb.2016.00986
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author Natarajan, Vengadesh Perumal
Zhang, Xinxu
Morono, Yuki
Inagaki, Fumio
Wang, Fengping
author_facet Natarajan, Vengadesh Perumal
Zhang, Xinxu
Morono, Yuki
Inagaki, Fumio
Wang, Fengping
author_sort Natarajan, Vengadesh Perumal
collection PubMed
description Recovering high quality genomic DNA from environmental samples is a crucial primary step to understand the genetic, metabolic, and evolutionary characteristics of microbial communities through molecular ecological approaches. However, it is often challenging because of the difficulty of effective cell lysis without fragmenting the genomic DNA. This work aims to improve the previous SDS-based DNA extraction methods for high-biomass seafloor samples, such as pelagic sediments and metal sulfide chimney, to obtain high quality and high molecular weight of the genomic DNA applicable for the subsequent molecular ecological analyses. In this regard, we standardized a modified SDS-based DNA extraction method (M-SDS), and its performance was then compared to those extracted by a recently developed hot-alkaline DNA extraction method (HA) and a commercial DNA extraction kit. Consequently, the M-SDS method resulted in higher DNA yield and cell lysis efficiency, lower DNA shearing, and higher diversity scores than other two methods, providing a comprehensive DNA assemblage of the microbial community on the seafloor depositional environment.
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spelling pubmed-49175422016-07-21 A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments Natarajan, Vengadesh Perumal Zhang, Xinxu Morono, Yuki Inagaki, Fumio Wang, Fengping Front Microbiol Microbiology Recovering high quality genomic DNA from environmental samples is a crucial primary step to understand the genetic, metabolic, and evolutionary characteristics of microbial communities through molecular ecological approaches. However, it is often challenging because of the difficulty of effective cell lysis without fragmenting the genomic DNA. This work aims to improve the previous SDS-based DNA extraction methods for high-biomass seafloor samples, such as pelagic sediments and metal sulfide chimney, to obtain high quality and high molecular weight of the genomic DNA applicable for the subsequent molecular ecological analyses. In this regard, we standardized a modified SDS-based DNA extraction method (M-SDS), and its performance was then compared to those extracted by a recently developed hot-alkaline DNA extraction method (HA) and a commercial DNA extraction kit. Consequently, the M-SDS method resulted in higher DNA yield and cell lysis efficiency, lower DNA shearing, and higher diversity scores than other two methods, providing a comprehensive DNA assemblage of the microbial community on the seafloor depositional environment. Frontiers Media S.A. 2016-06-23 /pmc/articles/PMC4917542/ /pubmed/27446026 http://dx.doi.org/10.3389/fmicb.2016.00986 Text en Copyright © 2016 Natarajan, Zhang, Morono, Inagaki and Wang. 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
Natarajan, Vengadesh Perumal
Zhang, Xinxu
Morono, Yuki
Inagaki, Fumio
Wang, Fengping
A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
title A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
title_full A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
title_fullStr A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
title_full_unstemmed A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
title_short A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
title_sort modified sds-based dna extraction method for high quality environmental dna from seafloor environments
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4917542/
https://www.ncbi.nlm.nih.gov/pubmed/27446026
http://dx.doi.org/10.3389/fmicb.2016.00986
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