ITS2 sequencing of Norway spruce wood-inhabiting fungi
PCR: mix of forward primers ITS3mix (Tedersoo et al., 2014) + ITS3Oo (Riit et al., 2016) in combination with reverse primer ITS4ngsUni (Tedersoo and Lindahl, 2016). Forward primers were tagged with one of the 12-base Golay indexes with at least four differences to any other index (Tedersoo et al., 2018). Clean-up: PCR Purification Kit (Favorgen Biotech. Corp., Vienna). Library preparation: TruSeq Nano DNA Library Prep kit. Sequencing: 2 x 300-bp paired-end mode and v600 cycle chemistry with PhiX control (Illumina, Inc., USA). R1 and R2 reads are joined.