ITS2 region of the fungal  ribosomal DNA sequencing of poplar rhizosphere in greenhouse: genotype PB25  in substrate WasteRock
Libraries for sequencing were prepared according to Illuminas 16S Metagenomic Sequencing Library Preparation guide, with the exception of using Qiagen HotStar Taq DNA polymerase for the first PCR (amplicon PCR). The template specific primers were (without the overhang adapter sequence): ITS9F (5-GAACGCAGCRAAIIGYGA-3) and ITS4R (5- TCCTCCGCTTATTGATATGC-3). The first PCR (amplicon PCR) was carried out for 40 cycles with annealing temperature of 50C. Diluted pooled samples were loaded on an Illumina MiSeq and sequenced using a 500-cycles MiSeq Reagent Kit v2.