title: MiSeq of mycobiome on alder leaves in a stream
design: DNA-extraction of 6 leaf discs (d=19mm) with FAST DNA Spin Kit for Soil, 1:100 dilution of extracts followed by 3-step PCR with pre-amplification (Primermix from Tedersoo et al. 2014, 2015), ligation of 9 barcodes, 5 indices, and illumina adapters. PCR products were purified, DNA conc. was measured, single products were equalized to 4nM and pooled. Sequencing of 4.4 pM using 600-cycle V3-chemistry.