16S rRNA gene seq rat intestinal sample
DNA was extracted (PowerLyzer? PowerSoil? DNA isolation kit, MoBio 12855-100) from approx 250 mg intestinal material according to the protocol provided by the manufacturer with minor modifications. The PCR amplification of the V3-region of the 16S rRNA gene was performed in a single PCR reactions including fusion primers with the Ion Torrent sequencing adaptors and barcode. (PBU 5?-A-adapter-TCAG-barcode-CCTACGGGAGGCAGCAG-3?) and reverse primer (PBR 5?-trP1-adapter-ATTACCGCGGCTGCTGG-3?) in 20  l total reaction volume. DNA quantity was measured using Qubit? dsDNA HS assay (InvitrogenTM, Q32851) and samples were pooled to obtain equimolar libraries containing up to 86 samples (randomized) in each library.