Optimized bacterial community characterization through full-length 16S rRNA gene sequencing utilizing MinION nanopore technology.

ABSTRACT:

Background

Accurate identification of bacterial communities is crucial for research applications, diagnostics, and clinical interventions. Although 16S ribosomal RNA (rRNA) gene sequencing is a widely employed technique for bacterial taxonomic classification, it often results in misclassified or unclassified bacterial taxa. This study sought to refine the full-length 16S rRNA gene sequencing protocol using the MinION sequencer, focusing on the V1-V9 regions. Our methodological enquiry examined several factors, including the number of PCR amplification cycles, choice of primers and Taq polymerase, and specific sequence databases and workflows employed. We used a microbial standard comprising eight bacterial strains (five gram-positive and three gram-negative) in known proportions as

SUBMITTER: Bertolo A 

PROVIDER: S-EPMC10870487 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature