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用于高通量测序微生物群落的 DNA/RNA 提取方法比较。

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

机构信息

Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Center for Microbiome Innovation, University of California, San Diego, La Jolla, CA, USA.

出版信息

Biotechniques. 2021 Mar;70(3):149-159. doi: 10.2144/btn-2020-0153. Epub 2021 Jan 29.

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.

摘要

微生物生态学研究人员的目标之一是从样本中的所有生物中获取最大量的信息。最近由 RNA 病毒 SARS-CoV-2 引起的 COVID-19 大流行凸显了传统基于 DNA 的方案的一个差距,包括作者之前建立的作为现场标准的高通量方法。为了能够同时进行 SARS-CoV-2 和微生物群落分析,作者比较了两种总核酸提取方案与作者之前经过基准测试的方案的相对性能。作者包括了一系列不同的环境和宿主相关样本类型,包括通常用于 COVID-19 检测的拭子取样部位。在这里,作者介绍了这些方案之间在成本、处理时间、DNA 和 RNA 产量、微生物群落组成、检测限和孔间污染方面的比较结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a973/7983038/d0b74072808e/btn-70-149-g1.jpg

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