Considerations and best practices for 16S rRNA gene sequencing studies

Graphical Abstract

The microbiome participates in numerous physiological activities that underlie metabolic health in mammals (Alvarez et al, 2020) with perturbation of the microbiome associated with disease in humans (So and Savidge, 2021). Several pre- and probiotics are consumed by both man and beast as a means of altering the gut microbiome for improved health and performance (Hughes and Holsher, 2021; van Wagenberg et al., 2020). The continued interest in livestock gut, fecal and intestinal microbiomes coupled with low cost DNA sequencing capabilities has enhanced our understanding of the complexities of bacterial communities that drive metabolic health. To assist with interpretation of microbiome profiling experiments, Weinroth et al. (2022) have compiled a suggested guide for best practices for microbiome profiling. The review provides an overview of 16S rRNA sequencing and considerations for performing large scale bacterial interrogations.