rRNA 16S of Capra hircus: semen
Semen from six adult goat bucks from the Murciano-Granadina breed (24 ejaculates, ##: 01 to 24) was used and refrigerated to 4 C using a skimmed milk-based extender, being stored at this temperature for a maximum of 24 h. Samples were taken at different steps of the preparation: in the crude ejaculates (ejaculates, E##), after the dilution with the refrigeration extender (diluted, D##), immediately after reaching 4 C (Chilled 0 h, R0##) and in samples refrigerated to 4 C and stored at this temperature for 24 h (Chilled 24 h, S##). Total DNA was extracted from 96 samples and PCR amplification of the bacterial 16S rRNA gene V3V4 regions was performed using forward and reverse primers. Libraries were normalized, purified and quantified on an ABI 7900HT real-time cycler. Sequencing was performed using the Illumina MiSeq platform with 2 x 300 bp reads.