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Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis

OBJECTIVE: Targeted sequencing of 16S rDNA amplicons is routinely used for microbial community profiling but this method suffers several limitations such as bias affinity of universal primers and short read size. Gene capture by hybridization represents a promising alternative. Here we used a metage...

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Autores principales: Cariou, Marie, Ribière, Céline, Morlière, Stéphanie, Gauthier, Jean-Pierre, Simon, Jean-Christophe, Peyret, Pierre, Charlat, Sylvain
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042230/
https://www.ncbi.nlm.nih.gov/pubmed/29996907
http://dx.doi.org/10.1186/s13104-018-3559-3
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author Cariou, Marie
Ribière, Céline
Morlière, Stéphanie
Gauthier, Jean-Pierre
Simon, Jean-Christophe
Peyret, Pierre
Charlat, Sylvain
author_facet Cariou, Marie
Ribière, Céline
Morlière, Stéphanie
Gauthier, Jean-Pierre
Simon, Jean-Christophe
Peyret, Pierre
Charlat, Sylvain
author_sort Cariou, Marie
collection PubMed
description OBJECTIVE: Targeted sequencing of 16S rDNA amplicons is routinely used for microbial community profiling but this method suffers several limitations such as bias affinity of universal primers and short read size. Gene capture by hybridization represents a promising alternative. Here we used a metagenomic extract from the pea aphid Acyrthosiphon pisum to compare the performances of two widely used PCR primer pairs with DNA capture, based on solution hybrid selection. RESULTS: All methods produced an exhaustive description of the 8 bacterial taxa known to be present in this sample. In addition, the methods yielded similar quantitative results, with the number of reads strongly correlating with quantitative PCR controls. Both methods can thus be considered as qualitatively and quantitatively robust on such a sample with low microbial complexity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-018-3559-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-60422302018-07-13 Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis Cariou, Marie Ribière, Céline Morlière, Stéphanie Gauthier, Jean-Pierre Simon, Jean-Christophe Peyret, Pierre Charlat, Sylvain BMC Res Notes Research Note OBJECTIVE: Targeted sequencing of 16S rDNA amplicons is routinely used for microbial community profiling but this method suffers several limitations such as bias affinity of universal primers and short read size. Gene capture by hybridization represents a promising alternative. Here we used a metagenomic extract from the pea aphid Acyrthosiphon pisum to compare the performances of two widely used PCR primer pairs with DNA capture, based on solution hybrid selection. RESULTS: All methods produced an exhaustive description of the 8 bacterial taxa known to be present in this sample. In addition, the methods yielded similar quantitative results, with the number of reads strongly correlating with quantitative PCR controls. Both methods can thus be considered as qualitatively and quantitatively robust on such a sample with low microbial complexity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-018-3559-3) contains supplementary material, which is available to authorized users. BioMed Central 2018-07-11 /pmc/articles/PMC6042230/ /pubmed/29996907 http://dx.doi.org/10.1186/s13104-018-3559-3 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Note
Cariou, Marie
Ribière, Céline
Morlière, Stéphanie
Gauthier, Jean-Pierre
Simon, Jean-Christophe
Peyret, Pierre
Charlat, Sylvain
Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
title Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
title_full Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
title_fullStr Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
title_full_unstemmed Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
title_short Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
title_sort comparing 16s rdna amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042230/
https://www.ncbi.nlm.nih.gov/pubmed/29996907
http://dx.doi.org/10.1186/s13104-018-3559-3
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