High-throughput amplicon sequencing of the full-length 16S rRNA gene with single-nucleotide resolution.

ABSTRACT: Targeted PCR amplification and high-throughput sequencing (amplicon sequencing) of 16S rRNA gene fragments is widely used to profile microbial communities. New long-read sequencing technologies can sequence the entire 16S rRNA gene, but higher error rates have limited their attractiveness when accuracy is important. Here we present a high-throughput amplicon sequencing methodology based on PacBio circular consensus sequencing and the DADA2 sample inference method that measures the full-length 16S rRNA gene with single-nucleotide resolution and a near-zero error rate. In two artificial communities of known composition, our method recovered the full complement of full-length 16S sequence variants from expected community members without residual errors. The measured abundances of intra-genomi

SUBMITTER: Callahan BJ 

PROVIDER: S-EPMC6765137 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature