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

微生物群落高通量测序的 DNA/RNA 提取方案比较

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作者: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 Mill

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.

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