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利用聚合酶链式反应分子指纹图谱方法和下一代测序技术的力量来理解微生物群落的结构和功能。

Harnessing the Power of PCR Molecular Fingerprinting Methods and Next Generation Sequencing for Understanding Structure and Function in Microbial Communities.

作者信息

Phadke Sujal, Salvador Andreia Filipa, Alves Joana Isabel, Bretschger Orianna, Alves Maria Madalena, Pereira Maria Alcina

机构信息

J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA, 92037, USA.

CEB-Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal.

出版信息

Methods Mol Biol. 2017;1620:225-248. doi: 10.1007/978-1-4939-7060-5_16.

Abstract

Polymerase chain reaction (PCR) is central to methods in molecular ecology. Here, we describe PCR-dependent approaches useful for investigating microbial diversity and its function in various natural, human-associated, and built environment ecosystems. Protocols routinely used for DNA extraction, purification, cloning, and sequencing are included along with various resources for the statistical analysis following gel electrophoresis-based methods (DGGE) and sequencing. We also provide insights into eukaryotic microbiome analysis, sample preservation techniques, PCR troubleshooting, DNA quantification methods, and commonly used ordination techniques.

摘要

聚合酶链反应(PCR)是分子生态学方法的核心。在此,我们描述了依赖PCR的方法,这些方法对于研究各种自然、人类相关和人造环境生态系统中的微生物多样性及其功能很有用。文中包括了常规用于DNA提取、纯化、克隆和测序的方案,以及基于凝胶电泳方法(DGGE)和测序后的统计分析的各种资源。我们还提供了关于真核生物微生物组分析、样品保存技术、PCR故障排除、DNA定量方法和常用排序技术的见解。

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