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A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities

One goal of microbial ecology researchers is to capture the maximum amount of information from all organisms in a sample. The recent COVID-19 pandemic, caused by the RNA virus SARS-CoV-2, has highlighted a gap in traditional DNA-based protocols, including the high-throughput methods the authors prev...

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Autores principales: Shaffer, Justin P, Marotz, Clarisse, Belda-Ferre, Pedro, Martino, Cameron, Wandro, Stephen, Estaki, Mehrbod, Salido, Rodolfo A, Carpenter, Carolina S, Zaramela, Livia S, Minich, Jeremiah J, Bryant, MacKenzie, Sanders, Karenina, Fraraccio, Serena, Ackermann, Gail, Humphrey, Gregory, Swafford, Austin D, Miller-Montgomery, Sandrine, Knight, Rob
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7931620/
https://www.ncbi.nlm.nih.gov/pubmed/33512248
http://dx.doi.org/10.2144/btn-2020-0153
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author Shaffer, Justin P
Marotz, Clarisse
Belda-Ferre, Pedro
Martino, Cameron
Wandro, Stephen
Estaki, Mehrbod
Salido, Rodolfo A
Carpenter, Carolina S
Zaramela, Livia S
Minich, Jeremiah J
Bryant, MacKenzie
Sanders, Karenina
Fraraccio, Serena
Ackermann, Gail
Humphrey, Gregory
Swafford, Austin D
Miller-Montgomery, Sandrine
Knight, Rob
author_facet Shaffer, Justin P
Marotz, Clarisse
Belda-Ferre, Pedro
Martino, Cameron
Wandro, Stephen
Estaki, Mehrbod
Salido, Rodolfo A
Carpenter, Carolina S
Zaramela, Livia S
Minich, Jeremiah J
Bryant, MacKenzie
Sanders, Karenina
Fraraccio, Serena
Ackermann, Gail
Humphrey, Gregory
Swafford, Austin D
Miller-Montgomery, Sandrine
Knight, Rob
author_sort Shaffer, Justin P
collection PubMed
description One goal of microbial ecology researchers is to capture the maximum amount of information from all organisms in a sample. The recent COVID-19 pandemic, caused by the RNA virus SARS-CoV-2, has highlighted a gap in traditional DNA-based protocols, including the high-throughput methods the authors previously established as field standards. To enable simultaneous SARS-CoV-2 and microbial community profiling, the authors compared the relative performance of two total nucleic acid extraction protocols with the authors' previously benchmarked protocol. The authors included a diverse panel of environmental and host-associated sample types, including body sites commonly swabbed for COVID-19 testing. Here the authors present results comparing the cost, processing time, DNA and RNA yield, microbial community composition, limit of detection and well-to-well contamination between these protocols.
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spelling pubmed-79316202021-03-04 A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities Shaffer, Justin P Marotz, Clarisse Belda-Ferre, Pedro Martino, Cameron Wandro, Stephen Estaki, Mehrbod Salido, Rodolfo A Carpenter, Carolina S Zaramela, Livia S Minich, Jeremiah J Bryant, MacKenzie Sanders, Karenina Fraraccio, Serena Ackermann, Gail Humphrey, Gregory Swafford, Austin D Miller-Montgomery, Sandrine Knight, Rob Biotechniques Reports One goal of microbial ecology researchers is to capture the maximum amount of information from all organisms in a sample. The recent COVID-19 pandemic, caused by the RNA virus SARS-CoV-2, has highlighted a gap in traditional DNA-based protocols, including the high-throughput methods the authors previously established as field standards. To enable simultaneous SARS-CoV-2 and microbial community profiling, the authors compared the relative performance of two total nucleic acid extraction protocols with the authors' previously benchmarked protocol. The authors included a diverse panel of environmental and host-associated sample types, including body sites commonly swabbed for COVID-19 testing. Here the authors present results comparing the cost, processing time, DNA and RNA yield, microbial community composition, limit of detection and well-to-well contamination between these protocols. Future Science Ltd 2021-01-29 2021-03 /pmc/articles/PMC7931620/ /pubmed/33512248 http://dx.doi.org/10.2144/btn-2020-0153 Text en © 2021 Justin P. Shaffer This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Reports
Shaffer, Justin P
Marotz, Clarisse
Belda-Ferre, Pedro
Martino, Cameron
Wandro, Stephen
Estaki, Mehrbod
Salido, Rodolfo A
Carpenter, Carolina S
Zaramela, Livia S
Minich, Jeremiah J
Bryant, MacKenzie
Sanders, Karenina
Fraraccio, Serena
Ackermann, Gail
Humphrey, Gregory
Swafford, Austin D
Miller-Montgomery, Sandrine
Knight, Rob
A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities
title A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities
title_full A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities
title_fullStr A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities
title_full_unstemmed A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities
title_short A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities
title_sort comparison of dna/rna extraction protocols for high-throughput sequencing of microbial communities
topic Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7931620/
https://www.ncbi.nlm.nih.gov/pubmed/33512248
http://dx.doi.org/10.2144/btn-2020-0153
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