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


Journal

BioTechniques
ISSN: 1940-9818
Titre abrégé: Biotechniques
Pays: England
ID NLM: 8306785

Informations de publication

Date de publication:
03 2021
Historique:
pubmed: 30 1 2021
medline: 30 3 2021
entrez: 29 1 2021
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 33512248
doi: 10.2144/btn-2020-0153
pmc: PMC7931620
doi:

Substances chimiques

DNA, Viral 0
RNA, Ribosomal, 16S 0

Types de publication

Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

149-159

Subventions

Organisme : NIGMS NIH HHS
ID : K12 GM068524
Pays : United States
Organisme : NIAID NIH HHS
ID : U01 AI124316
Pays : United States
Organisme : NCCIH NIH HHS
ID : DP1 AT010885
Pays : United States
Organisme : NIA NIH HHS
ID : U19 AG063744
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL134887
Pays : United States
Organisme : NIA NIH HHS
ID : RF1 AG058942
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK102932
Pays : United States
Organisme : NIH HHS
ID : 1RF1-AG058942-01,R01DK102932,R01HL134887,R01HL140976,U01AI124316,U19AG063744
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL140976
Pays : United States

Commentaires et corrections

Type : UpdateOf

Auteurs

Justin P Shaffer (JP)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Clarisse Marotz (C)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Pedro Belda-Ferre (P)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Cameron Martino (C)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.
Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.
Bioinformatics & Systems Biology Program, University of California San Diego, La Jolla, CA, USA.

Stephen Wandro (S)

Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.
Micronoma Inc., San Diego, CA, USA.

Mehrbod Estaki (M)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Rodolfo A Salido (RA)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Carolina S Carpenter (CS)

Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.

Livia S Zaramela (LS)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Jeremiah J Minich (JJ)

Marine Biology Research Division, University of California, San Diego, La Jolla, CA, USA.

MacKenzie Bryant (M)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Karenina Sanders (K)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Serena Fraraccio (S)

Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.
Micronoma Inc., San Diego, CA, USA.

Gail Ackermann (G)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Gregory Humphrey (G)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Austin D Swafford (AD)

Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.

Sandrine Miller-Montgomery (S)

Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.
Micronoma Inc., San Diego, CA, USA.

Rob Knight (R)

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.
Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.
Department of Computer Science & Engineering, University of California, San Diego, La Jolla, CA, USA.
Micronoma Inc., San Diego, CA, USA.

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Classifications MeSH