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

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.

Document Details

Document Type
Pub Defense Publication
Publication Date
Mar 01, 2021
Source ID
10.2144/btn-2020-0153

Entities

People

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

Organizations

  • Crohn's & Colitis Foundation of America
  • Defense Advanced Research Projects Agency
  • National Institutes of Health
  • United States Department of Defense
  • University of California, San Diego

Tags

Readers

  • Computational Modeling and Simulation
  • Microbial Pathology
  • Molecular Genetics

Technology Areas

  • Biotechnology