16s sequencing of mus musculus, female intestine mucosal scraping, Western Diet 10
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).