BioTechniques (Mar 2021)

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

  • Justin P Shaffer,
  • Clarisse Marotz,
  • Pedro Belda-Ferre,
  • Cameron Martino,
  • Stephen Wandro,
  • Mehrbod Estaki,
  • Rodolfo A Salido,
  • Carolina S Carpenter,
  • Livia S Zaramela,
  • Jeremiah J Minich,
  • MacKenzie Bryant,
  • Karenina Sanders,
  • Serena Fraraccio,
  • Gail Ackermann,
  • Gregory Humphrey,
  • Austin D Swafford,
  • Sandrine Miller-Montgomery,
  • Rob Knight

DOI
https://doi.org/10.2144/btn-2020-0153
Journal volume & issue
Vol. 70, no. 3
pp. 149 – 159

Abstract

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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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