Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis

The 16S rRNA gene has been a mainstay of sequence-based bacterial analysis for decades. However, high-throughput sequencing of the full gene has only recently become a realistic prospect. Here, we use in silico and sequence-based experiments to critically re-evaluate the potential of the 16S gene to provide taxonomic resolution at species and strain level. We demonstrate that targeting of 16S variable regions with short-read sequencing platforms cannot achieve the taxonomic resolution afforded by sequencing the entire (~1500 bp) gene. We further demonstrate that full-length sequencing platforms are sufficiently accurate to resolve subtle nucleotide substitutions (but not insertions/deletions) that exist…

• NI
  National Institutes of Health

  Award: 1U54DE02378901
• NI
  National Institute of Diabetes and Digestive and Kidney Diseases
• CF
  Common Fund