gut microbiota-16s
Microbial genomic DNA was extracted from fecal samples, and its quality was verified by 1% agarose gel electrophoresis. The hypervariable V3-V4 region of the 16S rRNA gene was amplified by PCR, and the resulting amplicons were purified using the Agencourt AMPure XP beads. Sequencing libraries were constructed and sequenced on an Illumina platform to generate paired-end reads. The raw sequencing data first underwent quality control using FastQC. Paired-end reads were merged, quality-filtered, and denoised, and chimeras were removed using QIIME2 to generate high-quality amplicon sequence variants (ASVs). These high-quality ASVs were used for downstream analyses, including alpha diversity (within-sample diversity), beta diversity (between-sample diversity), and taxonomic composition analysis.