16S rRNA gene amplicon of meat production microbiota: Sus domesticus tonsils
Palantine tonsils were removed just after evisceration, trimmed, cut into small pieces, and homogenized in 0.9% saline, ratio 1:5 (w/v), in stomacher bags. 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.