Mock 1 (CRX2193221)
Total genomic DNA was extracted from rat colonic samples using the QIAamp PowerFecal Pro DNA Kit. The hypervariable V3-V4 region of the bacterial 16S rRNA gene was amplified with universal primers 338F and 806R. PCR products were purified using AMPure XP beads. Subsequently, sequencing libraries were constructed through end-repair, adapter ligation, and index PCR. The final libraries were quantified, pooled in equimolar ratios, and sequenced on an Illumina NextSeq 2000 platform with a paired-end 2x151 bp strategy.