title: rumen microbial diversity
design: Fast DNA SPIN Kit for Soil (MP Bio, Biomedicals, Illkirch, France) extraction kit was used to extract metagenomic. A PCR was employed to amplify the V3 and V4 regions of the 16S ribosomal RNA gene 16S rRNA for bacteria. The PCR fragments were purified using the ZymocleanTM Gel DNA Recovery kit following the manufacturer's instructions. Composite samples for sequencing were created by combining equimolarratios of amplicons from the duplicate samples. Sequencing was performed using the Ion Torrent Personal Genome Machine (Life Technologies, Carlsbad, CA, USA) using theIon 314TM Chip Kit v2.