The Madness of Microbiome: Attempting To Find Consensus “Best Practice” for 16S Microbiome Studies

• Published in: Applied and environmental microbiology Vol. 84; no. 7; p. e02627-17
• Main Authors: Pollock, Jolinda, Glendinning, Laura, Wisedchanwet, Trong, Watson, Mick
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01.04.2018
• Subjects: Best practice; Community composition; Continuous improvement; Data analysis; Data processing; DNA Barcoding, Taxonomic - instrumentation; Gene expression; Gene sequencing; Genetic testing; Microbial activity; Microbiomes; Microbiota; Microbiota - genetics; Microorganisms; Minireview; Next-generation sequencing; Ribonucleic acid; RNA; RNA, Bacterial - analysis; RNA, Ribosomal, 16S - analysis; rRNA 16S; microbiome; 16S RNA; DNA sequencing
• ISSN: 0099-2240; 1098-5336; 1098-5336
• DOI: 10.1128/AEM.02627-17

The development and continuous improvement of high-throughput sequencing platforms have stimulated interest in the study of complex microbial communities. Currently, the most popular sequencing approach to study microbial community composition and dynamics is targeted 16S rRNA gene metabarcoding. To prepare samples for sequencing, there are a variety of processing steps, each with the potential to introduce bias at the data analysis stage. In this short review, key information from the literature pertaining to each processing step is described, and consequently, general recommendations for future 16S rRNA gene metabarcoding experiments are made.