16S amplicon sequencing of bos taurus: rumen fluid
Rumen fluid was collected through the oral stomach tube (OST) method (Muizelaar et al., 2020). Samples were snap frozen in dry ice and stored at -80 C until DNA extraction. DNA from rumen fluid samples was purified using the Qiagen DNeasy Blood and tissue Kit (Qiagen, Hilden, GERMANY). Sequencing libraries were prepared by a two-step polymerase chain reaction (PCR)-based workflow. The V4 region of the 16S rRNA gene was amplified using the region-specific primers V4-515F (GTGCCAGCMGCCGCGGTA) and V4-806R (GACTACHVGGGTATCTAATCC)