Evaluating the efficiency of 16S-ITS-23S operon sequencing for species level resolution in microbial communities.

ABSTRACT: Rapid advancements in long-read sequencing have facilitated species-level microbial profiling through full-length 16S rRNA sequencing (~ 1500 bp), and more notably, by the newer 16S-ITS-23S ribosomal RNA operon (RRN) sequencing (~ 4500 bp). RRN sequencing is emerging as a superior method for species resolution, exceeding the capabilities of short-read and full-length 16S rRNA sequencing. However, being in its early stages of development, RRN sequencing has several underexplored or understudied elements, highlighting the need for a critical and thorough examination of its methodologies. Key areas that require detailed analysis include understanding how primer pairs, sequencing platforms, and classifiers and databases affect the accuracy of species resolution achieved through RRN sequencing.

SUBMITTER: Srinivas M 

PROVIDER: S-EPMC11754871 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature