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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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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. |
format | Online Article Text |
id | pubmed-6042230 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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