Paired-end reads of gastrointestinal tract contents: adult male sheep: Sample Gastrointestinal tract contents of sheep
The V4 region of the 16S rRNA gene was amplified from genomic DNA using the primers (515F: 5’-GTGCCAGCMGCCGCGGTAA-3’, 806R: GGACTACHVGGGTWTCTAAT -3’) (Caporaso et al., 2011). The reverse primer is barcoded with a unique 6–bp error–correcting to each sample. Briefly, PCR reaction was performed in triplicate 20 µL containing  10 ng template DNA, 0.2 µM of each primer, 4 µL 5×FastPfu Buffer, 0.4 µL FastPfu Polymerase, and 2 µL 2.5 mM dNTPs (MBI Fermentas, Waltham, MA, USA). The following thermal cycling condition were used: 3 min initial denaturation at 94 °C for; 35 cycles of denaturation at 94 °C for 45 s, annealing at 50 °C for 60 s, and elongation at 72 °C for 90 s; and a last step at 72 °C for 10 min. 