16s sequencing of mus musculus, female intestine mucosal scraping, Western Diet + Empty Vector 6
DNA extraction and sequencing of the 16S ribosomal RNA gene was performed for mouse fecal samples as previously described in Jacobs, J et al (2017). In brief, bacterial DNA was extracted using the ZymoBIOMICS DNA kit (cat#D4300) with bead beating. The V4 region of the 16S gene was amplified and barcoded using 515f/806r primers then 250x2 bp sequencing was performed on an Illumina Miseq system. Raw data were processed using DADA2 scripts in R platform and quality-filtered reads (~115002 average reads per sample) were used to identify amplicon sequence variants (ASV) by closed reference picking against the Silva database (Callahan BJ et al, 2016).