16S rRNA gene amplicon of meat production microbiota: Sus domesticus skin
Swabs (~400cm2 area) were taken from the neck area, using sterile gauze pads (10x10 cm, 16 ply) moistened with 15 ml 0.9% saline, and homogenized in stomacher bags with 25 ml 0.9% saline. 10 ml swab homogenate was centrifuged for 20 min at 2,880 x g, and the pellets resuspended in 1 ml 0.9% saline. DNA extracted using MoBio PowerLyzer® Power Soil® DNA Isolation Kit modified by using a Mixer Mill MM 300 for 10 min at 30 cycles per second. DNA was diluted ten-fold and Two-step PCR of the v3-v4 region was performed, First 30 cycles with broad range primers (341F and 806R), then 15 cycles to add sequencing primers and adaptors. 250 bp Paired-end sequencing were performed on the Illumina MiSeq System using MiSeq Reagent Kits v2.