ZIEL - Institute for Food & Health, Technical University of Munich, 85354 Freising, Germany.
Chair of Nutrition and Immunology, Technical University of Munich, Gregor-Mendel-Str. 2, 85354 Freising, Germany.
STAR Protoc. 2020 Oct 26;1(3):100148. doi: 10.1016/j.xpro.2020.100148. eCollection 2020 Dec 18.
Targeted sequencing of 16S rRNA genes enables the analysis of microbiomes. Here, we describe a protocol for the collection, storage, and preparation of fecal samples. We describe how we cluster similar sequences and assign bacterial taxonomies. Using diversity analysis and machine learning, we can extract disease-associated features. We also describe a circadian analysis to identify the presence or absence of rhythms in taxonomies. Differences in rhythmicity between cohorts can contribute to determining disease-associated bacterial signatures. For complete details on the use and execution of this protocol, please refer to Reitmeier et al. (2020).
16S rRNA 基因靶向测序可用于分析微生物组。本实验描述了粪便样本的采集、储存和准备方法,包括如何对相似序列进行聚类并分配细菌分类群。通过多样性分析和机器学习,可以提取与疾病相关的特征。本实验还描述了一种节律分析方法,用于识别分类群中是否存在节律。不同队列间节律的差异有助于确定与疾病相关的细菌特征。有关本实验方案使用和执行的详细信息,请参考 Reitmeier 等人(2020 年)的文献。