16S rRNA gene amplicon of meat production microbiota: Sus domesticus feces
Colon content samlped directly from colon spiral were diluted 1:9 in 0.9% saline and vortexed, 1.5 ml centrifuged and 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.