Metagenome: JCF1
PCR amplification of fungal ribosomal internal transcribed spacers (region ITS2) was performed using primers ITS3/ITS4. 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).