fungal ITS2 amplicons from mycelia obtained from in-growth mesh bags
2-step PCR using with primers ITS4 and gITS7 (Ihrmark et al. 2012) containing partial TruSeq adapter sequences at their 5 end. The first PCR was done in two replicate 25 l reactions using Phusion Hot Start II polymerase with cycling conditions: an denaturation step at 98C for 30s, followed by 15 cycles at 98C for 10s, 55C for 30s, 72C for 10s, and an extension for 5 minutes.