Microbiome analysis via 16S and ITS amplicon sequencing is pivotal for studying microbial community composition and diversity, spanning bacteria and fungi. The 16S rRNA gene, found in bacteria and archaea, and ITS for fungi are key markers used for taxonomic classification.
The process entails sample collection, DNA extraction, PCR amplification targeting variable regions of 16S rRNA or ITS genes, library preparation, and high-throughput sequencing. Taxonomic classification involves clustering reads into operational taxonomic units (OTUs) or amplicon sequence variants (ASVs), followed by comparison to reference databases. Diversity and community analyses assess microbial community structure using metrics like alpha and beta diversity, unveiling richness, evenness, and composition insights.
Libraries are prepared using the QIAseq 16S/ITS Panels (Qiagen), developed for sequencing 16S rRNA and ITS regions on Illumina platforms and to perform a parallel profiling of bacterial and fungal communities.