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

Authors :
Shaffer JP
Marotz C
Belda-Ferre P
Martino C
Wandro S
Estaki M
Salido RA
Carpenter CS
Zaramela LS
Minich JJ
Bryant M
Sanders K
Fraraccio S
Ackermann G
Humphrey G
Swafford AD
Miller-Montgomery S
Knight R
Source :
BioTechniques [Biotechniques] 2021 Mar; Vol. 70 (3), pp. 149-159. Date of Electronic Publication: 2021 Jan 29.
Publication Year :
2021

Abstract

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.

Details

Language :
English
ISSN :
1940-9818
Volume :
70
Issue :
3
Database :
MEDLINE
Journal :
BioTechniques
Publication Type :
Academic Journal
Accession number :
33512248
Full Text :
https://doi.org/10.2144/btn-2020-0153