Metagenomic analysis of rumen microbiota; Ovis aries under high-grain diet
Total DNA was extracted from 200 L of rumen fluid using the DNeasy PowerSoil Pro Kit (Qiagen). The V3-V4 hypervariable regions of the 16S rRNA gene were amplified using 341F/806R primers. Libraries were constructed using the NEBNext Ultra II DNA Library Prep Kit with 8 PCR cycles. Fragment size selection (450-550 bp) was performed with AMPure XP beads. Library quality was assessed using Agilent 2100 Bioanalyzer.