Rapin Alexis, Pattaroni Céline, Marsland Benjamin J, Harris Nicola L
Global Health Institute, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
Faculty of Biology and Medicine, University of Lausanne, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland.
Curr Protoc Mouse Biol. 2017 Jun 19;7(2):100-129. doi: 10.1002/cpmo.29.
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.
微生物群已被证明在包括免疫、代谢和消化在内的多种生物过程中发挥重要作用。由于其成员的生长特性往往未知，评估微生物群的确切组成已被证明具有挑战性，基于培养的方法通常无法捕捉到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手段来收集有关微生物群中已培养和未培养成员的信息。本文提供了一种使用高通量测序来表征细菌群落物种组成的方法。简而言之，通过提取微生物群样本的全部DNA含量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约翰威立父子公司版权所有。
