Microbiota Analysis Using an Illumina MiSeq Platform to Sequence 16S rRNA Genes

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2017-06-19 DOI:10.1002/cpmo.29

Alexis Rapin, Céline Pattaroni, Benjamin J. Marsland, Nicola L. Harris

引用次数: 33

Abstract

The microbiota have been shown to play an important role in diverse biological processes including immunity, metabolism, and digestion. Assessing the exact composition of the microbiota has proven challenging due to the often unknown growth specificities of its members, and culture-based approaches typically fail to capture the complete diversity of microorganisms present. Next Generation Sequencing (NGS) methods provide an efficient means to gather information about cultured and uncultured members of the microbiota. This article provides a method to characterize bacterial communities in terms of species composition using high-throughput sequencing. Briefly, by extracting the entire DNA content of a microbiota sample and performing a targeted high-throughput sequencing of the 16S rRNA gene, a phylogenetic marker for prokaryotes, prediction of the composition of the entire bacterial community is made possible. © 2017 by John Wiley & Sons, Inc.

查看原文本刊更多论文

利用Illumina MiSeq平台对16S rRNA基因进行微生物群分析

微生物群已被证明在多种生物过程中发挥重要作用，包括免疫、代谢和消化。由于其成员的生长特异性通常未知，评估微生物群的确切组成已被证明具有挑战性，并且基于培养的方法通常无法捕获存在的微生物的完整多样性。下一代测序(NGS)方法提供了一种有效的方法来收集有关培养和未培养微生物群成员的信息。本文提供了一种方法来表征细菌群落方面的物种组成使用高通量测序。简而言之，通过提取微生物群样本的全部DNA内容，并对原核生物的系统发育标记16S rRNA基因进行靶向高通量测序，可以预测整个细菌群落的组成。©2017 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

自引率

0.00%

发文量

期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.