title: Illumina MiSeq sequencing; qiita_ptid_10724:13751.NGS18.0277.V3V4
design: High-throughput sequencing of the V3V4 region was performed by Eurofins on an Illumina MiSeq platform according to the standard protocols with minor adjustments. Amplicons were generated using a two-step PCR protocol. Briefly, the ITS2 region was PCR-amplified using universal primers that contained the adapter overhang nucleotide sequences for forward and reverse index primers. Amplicons were purified using AMPure XP beads (Beckman Coulter) and set up for the index PCR with Nextera XT index primers (Illumina). The indexed samples were purified using AMPure XP beads, quantified using the Fragment Analyzer (Advanced Analytical) and equal quantities from each sample were pooled.