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Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing

We are colonized by a vast population of genetically diverse microbes, the majority of which are unculturable bacteria that reside within the gastrointestinal tract. As affordable, advanced next-generation sequencing technologies become more widely available, important discoveries about the composit...

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Autores principales: McGaughey, Kara D., Yilmaz-Swenson, Tulay, Elsayed, Nourhan M., Cruz, Dianne A., Rodriguez, Ramona R., Kritzer, Michael D., Peterchev, Angel V., Gray, Megan, Lewis, Samantha R., Roach, Jeffrey, Wetsel, William C., Williamson, Douglas E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6107275/
https://www.ncbi.nlm.nih.gov/pubmed/30138447
http://dx.doi.org/10.1371/journal.pone.0202858
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author McGaughey, Kara D.
Yilmaz-Swenson, Tulay
Elsayed, Nourhan M.
Cruz, Dianne A.
Rodriguez, Ramona R.
Kritzer, Michael D.
Peterchev, Angel V.
Gray, Megan
Lewis, Samantha R.
Roach, Jeffrey
Wetsel, William C.
Williamson, Douglas E.
author_facet McGaughey, Kara D.
Yilmaz-Swenson, Tulay
Elsayed, Nourhan M.
Cruz, Dianne A.
Rodriguez, Ramona R.
Kritzer, Michael D.
Peterchev, Angel V.
Gray, Megan
Lewis, Samantha R.
Roach, Jeffrey
Wetsel, William C.
Williamson, Douglas E.
author_sort McGaughey, Kara D.
collection PubMed
description We are colonized by a vast population of genetically diverse microbes, the majority of which are unculturable bacteria that reside within the gastrointestinal tract. As affordable, advanced next-generation sequencing technologies become more widely available, important discoveries about the composition and function of these microbes become increasingly possible. In addition to rapid advancement in sequencing technologies, automated systems have been developed for nucleic acid extraction; however, these methods have yet to be widely used for the isolation of bacterial DNA from fecal samples. Here, we adapted Promega’s Maxwell(®) RSC PureFood GMO and Authentication kit for use with fecal samples and compared it to the commonly used Qiagen QIAamp(®) PowerFecal(®) kit. Results showed that the two approaches yielded similar measures of DNA purity and successful next-generation sequencing amplification and produced comparable composition of microbial communities. However, DNA extraction with the Maxwell(®) RSC kit produced higher concentrations with a lower fecal sample input weight and took a fraction of the time compared to the QIAamp(®) PowerFecal(®) protocol. The results of this study demonstrate that the Promega Maxwell(®) RSC system can be used for medium-throughput DNA extraction in a time-efficient manner without compromising the quality of the downstream sequencing.
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spelling pubmed-61072752018-08-30 Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing McGaughey, Kara D. Yilmaz-Swenson, Tulay Elsayed, Nourhan M. Cruz, Dianne A. Rodriguez, Ramona R. Kritzer, Michael D. Peterchev, Angel V. Gray, Megan Lewis, Samantha R. Roach, Jeffrey Wetsel, William C. Williamson, Douglas E. PLoS One Research Article We are colonized by a vast population of genetically diverse microbes, the majority of which are unculturable bacteria that reside within the gastrointestinal tract. As affordable, advanced next-generation sequencing technologies become more widely available, important discoveries about the composition and function of these microbes become increasingly possible. In addition to rapid advancement in sequencing technologies, automated systems have been developed for nucleic acid extraction; however, these methods have yet to be widely used for the isolation of bacterial DNA from fecal samples. Here, we adapted Promega’s Maxwell(®) RSC PureFood GMO and Authentication kit for use with fecal samples and compared it to the commonly used Qiagen QIAamp(®) PowerFecal(®) kit. Results showed that the two approaches yielded similar measures of DNA purity and successful next-generation sequencing amplification and produced comparable composition of microbial communities. However, DNA extraction with the Maxwell(®) RSC kit produced higher concentrations with a lower fecal sample input weight and took a fraction of the time compared to the QIAamp(®) PowerFecal(®) protocol. The results of this study demonstrate that the Promega Maxwell(®) RSC system can be used for medium-throughput DNA extraction in a time-efficient manner without compromising the quality of the downstream sequencing. Public Library of Science 2018-08-23 /pmc/articles/PMC6107275/ /pubmed/30138447 http://dx.doi.org/10.1371/journal.pone.0202858 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 (https://creativecommons.org/publicdomain/zero/1.0/) public domain dedication.
spellingShingle Research Article
McGaughey, Kara D.
Yilmaz-Swenson, Tulay
Elsayed, Nourhan M.
Cruz, Dianne A.
Rodriguez, Ramona R.
Kritzer, Michael D.
Peterchev, Angel V.
Gray, Megan
Lewis, Samantha R.
Roach, Jeffrey
Wetsel, William C.
Williamson, Douglas E.
Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing
title Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing
title_full Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing
title_fullStr Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing
title_full_unstemmed Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing
title_short Comparative evaluation of a new magnetic bead-based DNA extraction method from fecal samples for downstream next-generation 16S rRNA gene sequencing
title_sort comparative evaluation of a new magnetic bead-based dna extraction method from fecal samples for downstream next-generation 16s rrna gene sequencing
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6107275/
https://www.ncbi.nlm.nih.gov/pubmed/30138447
http://dx.doi.org/10.1371/journal.pone.0202858
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