Metabarcoding ITS2 region of fungi: peat sample from raised bog
The library was sequenced on Illumina MiSeq (length of reads  300 bp on both sides fragments) using MiSeq Reagent Kit v3 (600 cycles). FASTQ files were obtained using bcl2fastq v2.17.1.14 Conversion Software (Illumina). The PhiX phage library was used to control sequencing parameters. Most of the readings related to phage DNA was removed during demultiplexing. The samples of extracted DNA were sent for sequencing to outsource company (Evrogen, Moscow, https://evrogen.ru/). The quality of the obtained metagenomic DNA was checked by electrophoresis in agarose gel. Quantification was carried out by measuring the concentration of DNA by Qubit 2, using dsDNA HS reagent kit (ThermoFisher Scientific). Preparation of libraries for sequencing was carried out in accordance with the protocol described in 16S Metagenomic Sequencing Library Preparation (Part # 15044223 Rev. B; Illumina). Amplification of ITS variable regions was carried out using primers: Next-fITS7_For: 5'-GTGARTCATCGAATCTTTG-3' and Next-ITS4_Rev: 5'-TCCTCCGCTTATTGATATGC-3'. After obtaining the amplicons, the libraries were purified and pooled equimolarly with SequalPrep Normalization Plate Kit (ThermoFisher, Cat # A10510-01). Quality control of libraries was carried out using the Fragment Analyzer, and quantitative analysis was carried out with qPCR.