title: Amplicon-seq_Miseq_V3 of grower male pigs
design: Library construction of the V3 hypervariable region (from 16S rRNA gene) followed by sequencing on an Illumina MiSeq platform (paired end reads; 2*300 bp) were performed. Amplicons of the V3 hypervariable region of the 16S rRNA gene, were generated using the primer set V3_F (CCTACGGGAGGCAGCAG) and reverse primer V3_R (ATTACCGCGGCTGCTGG). Polymerase chain reaction conditions were as follows: 2 min at 98 C, 15 (10 s at 98 C, 30 s at 55 C, and 10 s at 72 C), and 7 min at 72 C. The PCR products were mixed with the same volume of 1x loading buffer (contained SYBR green) and were detected by electrophoresis on 2% agarose gel. Prior to library preparation, the PCR products were mixed in equimolar ratio and purified using Qiagen Gel Extraction Kit (Qiagen, Hilden, Germany).