title: Amplicon-MiSeq of metagenome: juvenile male rats cecum
design: Specimens were placed into a MoBio PowerMag Soil DNA Isolation Bead Plate. DNA was extracted following MoBios instructions on a KingFisher robot. Bacterial 16S rRNA genes were PCR-amplified with dual-barcoded primers targeting the V4 region (515F 5-GTGCCAGCMGCCGCGGTAA-3, and 806R 5-GGACTACHVGGGTWTCTAAT-3), as per the protocol of Kozich et al. (2013). Amplicons were sequenced with an Illumina MiSeq using the 300-bp paired-end kit (v.3). Sequences were denoised, taxonomically classified using Silva (v. 138) as the reference database.