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A comparison of DNA/RNA extraction protocols for high-throughput sequencing of microbial communities.
- 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.
- Subjects :
- Animals
Biodiversity
Cats
Chemical Fractionation methods
Feces microbiology
Feces virology
Female
Fermented Foods microbiology
Humans
Limit of Detection
Male
Metagenomics methods
Mice
Saliva microbiology
Saliva virology
Skin microbiology
Skin virology
DNA, Viral isolation & purification
High-Throughput Nucleotide Sequencing methods
Microbiota genetics
RNA, Ribosomal, 16S isolation & purification
SARS-CoV-2 genetics
Subjects
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