2.6. Microbiome Analyses
Fecal samples were collected and homogenized in OMNIgene GUT OM-200 tubes (DNA Genotek, ON, Canada) and transported to the laboratory at 4 °C. The samples were vortexed and aliquoted for storage at −80 °C. Whole genomic DNA was extracted using the Promega Fecal extraction kit with beads beating, and agarose gel electrophoresis was performed as quality control for the DNA samples. The DNA samples were subject to PCR amplification for the 16S rRNA gene V3–V4 region. The amplicons were purified and subject to library preparation and quantification followed by paired-end 250bp sequencing (100K tags per sample) on the NovaSeq (Illumina, San Diego, CA, USA). DADA2 [