Original title: Pipeline for species-resolved full-length16S rRNA amplicon nanopore sequencing analysis of low-complexity bacterial microbiota
Authors: Disha Tandon,Yuan Dong,Siegfried Hapfelmeier
This article discusses the use of 16S rRNA amplicon sequencing, a common method for studying prokaryotic microbial communities. While this technique is effective for examining complex microbiomes, it has limitations in terms of taxonomic classification. The authors present a new approach using third-generation sequencing technologies, specifically nanopore sequencing, which allows for full-length 16S rRNA gene sequencing. This enables greater taxonomic resolution, down to the species and strain levels. The researchers developed a pipeline that includes library preparation and computational analysis, and they applied it to a study of intestinal bacteria in mice. They found that the standard primers provided by Oxford Nanopore Technologies did not efficiently amplify the gene of interest, so they designed their own primers and used the company’s barcoding kit for library preparation. Their method provided accurate species-level classification and outperformed Illumina short-read sequencing, making it a cost-effective and accessible option for studying low-complexity microbiota.
Original article: https://www.biorxiv.org/content/10.1101/2023.12.05.570138v1