16S rRNA of rumen content: sheep
Genomic DNA of the rumen content microbiota was extracted using a DNA Kit (EZNA, Omega Bio-Tek, Norcross, GA) according to the manufacturers protocols. The quantity and quality of the microbial DNA were examined using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific) and 1.0% agarose gel electrophoresis. All extracted DNA samples were stored at -80C until subsequent processing. For the analysis of bacterial abundance and diversity, we used universal primers (341F: 5-CCTAYGGGRBGCASCAG-3, 806R: 5-GGACTACNNGGGTATCTAAT-3) to amplify the V3 and V4 regions of the 16S rRNA gene and with a 6 bp barcode unique to each sample. After PCR amplification, the Illumina MiSeq PE 312 platform was used to perform the sequencing of all amplicon libraries