Microbiota Analysis Using Sequencing by Synthesis: From Library Preparation to Sequencing

Nelson, A; Stewart, CJ

doi:10.1007/978-1-0716-0338-3_15

Microbiota Analysis Using Sequencing by Synthesis: From Library Preparation to Sequencing

Lookup NU author(s): Professor Christopher Stewart ORCiD

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This is the authors' accepted manuscript of a book chapter that has been published in its final definitive form by Humana Press Inc., 2020.

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Abstract

© 2020, Springer Science+Business Media, LLC, part of Springer Nature.The highly parallel nature of sequencing by synthesis (SBS) allows millions of amplicons to be sequenced simultaneously, which has led to enormous interest in the investigation of bacterial communities (often referred to as the microbiota). In this protocol, we describe a method for the ‘universal’ amplification of the v4 region of the bacterial 16S rRNA gene from genomic DNA and prepare these amplicons so that they can be sequenced using the MiSeq system (Illumina). The protocol provides instruction on sequencing of 188 genomic DNA samples plus PCR positive and negative controls, which can be applied to any sample type where bacterial DNA may be of interest.