Mouse Fecal Samples
DNA was extracted using the ZymoBIOMICS-96 MagBead DNA Kit (Zymo Research) on an automated platform. The V3-V4 region of the bacterial 16S rRNA gene was amplified using the Quick-16S NGS Library Prep Kit (Zymo Research) with custom primers designed for optimal coverage and sensitivity. PCR reactions were performed in real-time PCR machines to control cycles and limit chimera formation. The final pooled library was cleaned, quantified, and sequenced on an Illumina Nextseq with a P1 reagent kit (600 cycles) using a 30% PhiX spike-in.