Amplicon-Seq of rhizosphere metagenome: rice plant
PCR amplification of partial archaeal small-subunit ribosomal RNA genes (region V4V5 of 16S) was performed using primer 787F/1059R. 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).