16S V4 amplicon  of bovine metagenome: rumen
On day 20, four hours after feeding, 50 mL of ruminal content was collected, frozen at 80C, and used for DNA extraction by mechanical disruption and phenol method. DNA was quantified (Qubit, Invitrogen), adjusted to 10 ng/L, and amplified by PCR targeting the V4 region of the 16S rRNA gene using universal primers. Amplicons (~380 bp) were visualized in agarose gel, purified, and sequenced on a MiSeq platform (Illumina) with 5% PhiX control.
SUB15685777
B13P3_S13