16S amplicon sequencing of Mus musculus: adult female fecal sample
Mouse fecal samples were collected and stored at 80 C until processing. Genomic DNA was extracted using the QIAamp PowerFecal Pro DNA Kit (Qiagen) according to the manufacturers instructions. Microbial community composition was analyzed using two-stage 16S rRNA gene amplicon sequencing targeting the V1V3 regions, following methods previously described (PMID: 40763731, PMID: 29767361, PMID: 38188172). Briefly, the V1V3 region of the bacterial 16S rRNA gene was amplified using a mixture of 27F forward primers and the 534R reverse primer. Amplicons were generated using a two-step PCR protocol with Fluidigm common sequence linkers and Illumina adapter barcodes. Libraries were pooled and sequenced on an Illumina MiSeq platform using paired-end 2  300 bp reads.