Self-assembled community
Library preparation was performed following the Illumina 16S Metagenomic Sequencing Library Preparation protocol. Briefly, target regions were amplified using gene-specific primers with Illumina overhang adapters. Amplicons were purified using AMPure XP magnetic beads and subsequently subjected to a second PCR to attach dual indices and Illumina sequencing adapters. The final libraries were purified, quantified using a Qubit fluorometer, normalized, pooled, and denatured before loading onto the MiSeq instrument for paired-end sequencing.