Cargando…
Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing
The gastrointestinal tract contains a vast community of microbes that to this day remain largely unculturable, making studies in this area challenging. With the newly affordable advanced sequencing technology, important breakthroughs in this exciting field are now possible. However, standardized met...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4657925/ https://www.ncbi.nlm.nih.gov/pubmed/26599606 http://dx.doi.org/10.1371/journal.pone.0143334 |
_version_ | 1782402438493372416 |
---|---|
author | Hart, Marcia L. Meyer, Alexandra Johnson, Philip J. Ericsson, Aaron C. |
author_facet | Hart, Marcia L. Meyer, Alexandra Johnson, Philip J. Ericsson, Aaron C. |
author_sort | Hart, Marcia L. |
collection | PubMed |
description | The gastrointestinal tract contains a vast community of microbes that to this day remain largely unculturable, making studies in this area challenging. With the newly affordable advanced sequencing technology, important breakthroughs in this exciting field are now possible. However, standardized methods of sample collection, handling, and DNA extraction have yet to be determined. To help address this, we investigated the use of 5 common DNA extraction methods on fecal samples from 5 different species. Our data show that the method of DNA extraction impacts DNA concentration and purity, successful NGS amplification, and influences microbial communities seen in NGS output dependent on the species of fecal sample and the DNA extraction method used. These data highlight the importance of careful consideration of DNA extraction method used when designing and interpreting data from cross species studies. |
format | Online Article Text |
id | pubmed-4657925 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-46579252015-12-02 Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing Hart, Marcia L. Meyer, Alexandra Johnson, Philip J. Ericsson, Aaron C. PLoS One Research Article The gastrointestinal tract contains a vast community of microbes that to this day remain largely unculturable, making studies in this area challenging. With the newly affordable advanced sequencing technology, important breakthroughs in this exciting field are now possible. However, standardized methods of sample collection, handling, and DNA extraction have yet to be determined. To help address this, we investigated the use of 5 common DNA extraction methods on fecal samples from 5 different species. Our data show that the method of DNA extraction impacts DNA concentration and purity, successful NGS amplification, and influences microbial communities seen in NGS output dependent on the species of fecal sample and the DNA extraction method used. These data highlight the importance of careful consideration of DNA extraction method used when designing and interpreting data from cross species studies. Public Library of Science 2015-11-24 /pmc/articles/PMC4657925/ /pubmed/26599606 http://dx.doi.org/10.1371/journal.pone.0143334 Text en © 2015 Hart et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Hart, Marcia L. Meyer, Alexandra Johnson, Philip J. Ericsson, Aaron C. Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing |
title | Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing |
title_full | Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing |
title_fullStr | Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing |
title_full_unstemmed | Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing |
title_short | Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing |
title_sort | comparative evaluation of dna extraction methods from feces of multiple host species for downstream next-generation sequencing |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4657925/ https://www.ncbi.nlm.nih.gov/pubmed/26599606 http://dx.doi.org/10.1371/journal.pone.0143334 |
work_keys_str_mv | AT hartmarcial comparativeevaluationofdnaextractionmethodsfromfecesofmultiplehostspeciesfordownstreamnextgenerationsequencing AT meyeralexandra comparativeevaluationofdnaextractionmethodsfromfecesofmultiplehostspeciesfordownstreamnextgenerationsequencing AT johnsonphilipj comparativeevaluationofdnaextractionmethodsfromfecesofmultiplehostspeciesfordownstreamnextgenerationsequencing AT ericssonaaronc comparativeevaluationofdnaextractionmethodsfromfecesofmultiplehostspeciesfordownstreamnextgenerationsequencing |