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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...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Blackwell Publishing Ltd
2012
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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. |
format | Online Article Text |
id | pubmed-3815280 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Blackwell Publishing Ltd |
record_format | MEDLINE/PubMed |
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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