Sequencing of fermentation samples after 0.5 hours of rumen fluid in vitro
As described in the manufacturers instructions, total microbial genomic DNA from total incubation fluid was extracted by the FastDNA  SPIN for soil kit (MP Biomedicals, Solon, USA). DNA concentration and quality were evaluated by a NanoDrop ND-2000 spectrophotometer (Thermo Scientific Inc., USA) and 1.0% agarose gel electrophoresis. Polymerase chain reaction (PCR) was used to amplify the V3-V4 region of 16S rRNA gene by pair of primers 338F (5- ACTCCTACGGGAGGCAGCAG-3') and 806R (5'-GGACTACHVGGGTWTCTAAT-3')