Metagenome: JS1
PCR amplification of partial bacterial small-subunit ribosomal RNA genes (region V3V4 of 16S) was performed using primer 341F/805R. Both primers were tagged with Illumina adapter sequences while the reverse primer was barcoded to facilitate multiplexing. Each sample was amplified in triplicates, analyzed using 1.5% agarose gel, and amplicons from the triplicate PCRs were pooled and purified using Qiaquick gel extraction kit (Qiagen). The amplicons were mixed in equimolar concentrations and sequenced on the MiSeq platform (Illumina Inc., San Diego, CA, USA) with paired-end strategy (600 cycles v3).