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Biotechniques ; 70(3): 149-159, 2021 03.
Article in English | MEDLINE | ID: covidwho-1054921

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.


Subject(s)
DNA, Viral/isolation & purification , High-Throughput Nucleotide Sequencing/methods , Microbiota/genetics , RNA, Ribosomal, 16S/isolation & purification , SARS-CoV-2/genetics , 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
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