Bacteria in rumen and feces of cow on RFDDGS
Three grams (wet weight) of homogenized samples of the rumen fluid and feces from each cow were used for the DNA extraction with a bead-beating method followed by phenol-chloroform extraction. The DNA was quantified using a Nanodrop spectrophotometer following staining using a Quant-it Pico Green dsDNA kit (Invitrogen, Paisley, UK). The DNA samples were stored at -80C until further processing. The V3V4 region of the bacteria 16S ribosomal RNA gene was amplified by PCR (95C for 2 min, followed by 25 cycles at 95C for 30 s, 55C for 30 s and 72C of 30 s with a final extension at 72C for 5 min).