Amplicon of EKO 5
Metagenomic DNA was extracted using Zymo Researchs Quick-16S kit, followed by 16S ribosomal RNA gene sequencing targeting the V3-V4 region to identify microbial communities. Quality control and adapter trimming were performed with bcl-convert, and sequences were processed in QIIME2. Primer sequences were removed using QIIME2 cutadapt, and denoising was performed using QIIME2 DADA2. Operational taxonomic units (OTUs) were assigned using the Silva database and VSEARCH utility within QIIME2 feature-classifier. OTUs were collapsed to their lowest taxonomic units, and relative frequencies were calculated. Alpha and beta diversity analyses were conducted using QIIME2 q2-diversity to assess intra- and inter-sample variability.