title: Illumina HiSeq 2500 paired end sequencing; 16S sequencing of the stomach fluid of NG-8307_18_c
library_construction_protocol: DNA extraction, sequencing library construction and sequencing were conducted. Rumen samples were defrosted at 4°C overnight and vortexed until homogenous. A representative sample (500 µl) containing both rumen liquid and solids was used for DNA isolation. Two set of primers were used to create 16S rRNA libraries, one set for all bacteria and one set for all archaea. Universal bacterial 16S rRNA gene primers (covering the V1-V3 variable regions) 27F:5'-AGAGTTTGATCCTGGCTCAG-3' and 534R: 5'-ATTACCGCGGCTGCTGG-3' were used to generate the bacterial amplicon libraries (expected amplicon size 508 bp). Universal archaeal 16S rRNA gene primers (covering the V4-V6 variable regions) S-D-Arch-0519-a-S-15"5'-CAGCMGCCGCGGTAA-3' and S-D-Arch-1041-a-A-18:5'-GGCCATGCACCWCCTCTC-3' were used to generate the archaeal amplicon libraries (expected amplicon size 542 bp).