title: intestinal bacterial microbiota of bovine with and without tuberculosis, based on massive sequencing of amplicons from the V4 region of the 16S rRNA gene.
design: Fecal samples were collected aseptically of the intestinal segments. DNA extraction was performed using Precellys (Bertin Technologies) lysis for homogenization, following Mag Max Multi sample kit (Applied Biosystems) instructions. Libraries were generated by amplifying the V4 region of the bacterial 16S rRNA gene from bovine feces samples, using 515F and 806R primers containing the Illumina adapter sequences. Amplification was performed on the ProFlex PCR System (Life Technologies Inc, USA). The amplicons were checked by agarose gel electrophoresis and purified using Agencourt AMPure XP beads (Beckman Coulter, Indianapolis, Indiana, USA). Indexes were added following the protocol established by the manufacturer (Illumina Inc., San Diego, California, USA). After indexing, the samples underwent a final purification with Agencourt AMPure XP beads. Libraries were then quantified with Qubit and normalized to 8 pM. Sequencing was conducted on the Illumina MiSeq System with nano paired-end kit v2 500-cycles.
identifiers.namespace: SUB15167298
identifiers.id: 314S