Cecum, 12 weeks, replicate 1
Bacterial genomic DNA was extracted from approximately 100 mg of mouse small intestinal contents, cecal contents, or feces using the TIANamp Stool DNA Kit according to the manufacturers instructions. The V4 region of the bacterial 16S rRNA gene was amplified by PCR using primers 515F and 806R. PCR products of the expected size were verified and purified by 2% agarose gel electrophoresis. Purified amplicons from each sample were quantified, pooled in equal amounts, and used for library construction by end repair, A-tailing, and ligation of Illumina sequencing adapters. Libraries were sequenced on an Illumina platform using a 250 bp paired-end strategy.