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

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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.

Original languageEnglish
Pages (from-to)100-129
Number of pages30
JournalCurrent Protocols in Mouse Biology
Volume7
Issue number2
DOIs
Publication statusPublished - 19 Jun 2017
Externally publishedYes

Keywords

  • 16S rRNA
  • microbiota
  • MiSeq
  • next generation sequencing

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