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Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure

Three soil DNA extraction procedures (homemade protocols and commercial kit) varying in their practicability were applied to contrasting soils to evaluate their efficiency in recovering: (i) soil DNA and (ii) bacterial diversity estimated by 16S rDNA pyrosequencing. Significant differences in DNA yi...

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Autores principales: Terrat, Sébastien, Christen, Richard, Dequiedt, Samuel, Lelièvre, Mélanie, Nowak, Virginie, Regnier, Tiffanie, Bachar, Dipankar, Plassart, Pierre, Wincker, Patrick, Jolivet, Claudy, Bispo, Antonio, Lemanceau, Philippe, Maron, Pierre‐Alain, Mougel, Christophe, Ranjard, Lionel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3815280/
https://www.ncbi.nlm.nih.gov/pubmed/21989224
http://dx.doi.org/10.1111/j.1751-7915.2011.00307.x
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author Terrat, Sébastien
Christen, Richard
Dequiedt, Samuel
Lelièvre, Mélanie
Nowak, Virginie
Regnier, Tiffanie
Bachar, Dipankar
Plassart, Pierre
Wincker, Patrick
Jolivet, Claudy
Bispo, Antonio
Lemanceau, Philippe
Maron, Pierre‐Alain
Mougel, Christophe
Ranjard, Lionel
author_facet Terrat, Sébastien
Christen, Richard
Dequiedt, Samuel
Lelièvre, Mélanie
Nowak, Virginie
Regnier, Tiffanie
Bachar, Dipankar
Plassart, Pierre
Wincker, Patrick
Jolivet, Claudy
Bispo, Antonio
Lemanceau, Philippe
Maron, Pierre‐Alain
Mougel, Christophe
Ranjard, Lionel
author_sort Terrat, Sébastien
collection PubMed
description Three soil DNA extraction procedures (homemade protocols and commercial kit) varying in their practicability were applied to contrasting soils to evaluate their efficiency in recovering: (i) soil DNA and (ii) bacterial diversity estimated by 16S rDNA pyrosequencing. Significant differences in DNA yield were systematically observed between tested procedures. For certain soils, 10 times more DNA was recovered with one protocol than with the others. About 15 000 sequences of 16S rDNA were obtained for each sample which were clustered to draw rarefaction curves. These curves, as well as the PCA ordination of community composition based on OTU clustering, did not reveal any significant difference between procedures. Nevertheless, significant differences between procedures were highlighted by the taxonomic identification of sequences obtained at the phylum to genus levels. Depending on the soil, differences in the number of genera detected ranged from 1% to 26% between the most and least efficient procedures, mainly due to a poorer capacity to recover populations belonging to Actinobacteria, Firmicutes or Crenarchaeota. This study enabled us to rank the relative efficiencies of protocols for their recovery of soil molecular microbial biomass and bacterial diversity and to help choosing an appropriate soil DNA extraction procedure adapted to novel sequencing technologies.
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spelling pubmed-38152802014-02-12 Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure Terrat, Sébastien Christen, Richard Dequiedt, Samuel Lelièvre, Mélanie Nowak, Virginie Regnier, Tiffanie Bachar, Dipankar Plassart, Pierre Wincker, Patrick Jolivet, Claudy Bispo, Antonio Lemanceau, Philippe Maron, Pierre‐Alain Mougel, Christophe Ranjard, Lionel Microb Biotechnol Brief Reports Three soil DNA extraction procedures (homemade protocols and commercial kit) varying in their practicability were applied to contrasting soils to evaluate their efficiency in recovering: (i) soil DNA and (ii) bacterial diversity estimated by 16S rDNA pyrosequencing. Significant differences in DNA yield were systematically observed between tested procedures. For certain soils, 10 times more DNA was recovered with one protocol than with the others. About 15 000 sequences of 16S rDNA were obtained for each sample which were clustered to draw rarefaction curves. These curves, as well as the PCA ordination of community composition based on OTU clustering, did not reveal any significant difference between procedures. Nevertheless, significant differences between procedures were highlighted by the taxonomic identification of sequences obtained at the phylum to genus levels. Depending on the soil, differences in the number of genera detected ranged from 1% to 26% between the most and least efficient procedures, mainly due to a poorer capacity to recover populations belonging to Actinobacteria, Firmicutes or Crenarchaeota. This study enabled us to rank the relative efficiencies of protocols for their recovery of soil molecular microbial biomass and bacterial diversity and to help choosing an appropriate soil DNA extraction procedure adapted to novel sequencing technologies. Blackwell Publishing Ltd 2012-01 2011-12-14 /pmc/articles/PMC3815280/ /pubmed/21989224 http://dx.doi.org/10.1111/j.1751-7915.2011.00307.x Text en Copyright © 2011 The Authors. Microbial Biotechnology © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd
spellingShingle Brief Reports
Terrat, Sébastien
Christen, Richard
Dequiedt, Samuel
Lelièvre, Mélanie
Nowak, Virginie
Regnier, Tiffanie
Bachar, Dipankar
Plassart, Pierre
Wincker, Patrick
Jolivet, Claudy
Bispo, Antonio
Lemanceau, Philippe
Maron, Pierre‐Alain
Mougel, Christophe
Ranjard, Lionel
Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure
title Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure
title_full Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure
title_fullStr Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure
title_full_unstemmed Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure
title_short Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure
title_sort molecular biomass and metataxogenomic assessment of soil microbial communities as influenced by soil dna extraction procedure
topic Brief Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3815280/
https://www.ncbi.nlm.nih.gov/pubmed/21989224
http://dx.doi.org/10.1111/j.1751-7915.2011.00307.x
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