title: Fungal ITS2, isolated from root tissue collected from a field-grown Eucalyptus seedling
design: (Reference list can be found at https://doi.org/10.1111/mec.14536) DNA was extracted from each sample using a PowerSoil DNA isolation kit (MO BIO, Carlsbad, USA) following the manufacturer's instructions, except DNA was eluted with 50 ul of autoclaved milliQ water. An additional step involving sample disruption was conducted prior to the extraction, using 5 mm stainless steel beads for 30 s (soil), 1 min (roots) or 3 min (litter) at 30 Hz in a TissueLyser (Qiagen, Germany). Extracted DNA was quantified using a NanoDrop 2000/2000c Spectrophotometer (Thermo Scientific, Wilmington, USA), and then diluted with PCR-grade water to 5 ng/uL in 20uL. The ITS2 region was targeted for sequencing using fITS7 (5'-GTGARTCATCGAATCTTTG-3'; Ihrmark et al. 2012) and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'; White et al. 1990). Genomic libraries were prepared using the Nextera XT Index Kit (Illumina, San Diego, CA, USA) and paired-end (2 x 251 bases) sequencing was performed on the Illumina MiSeq platform at the Ramaciotti Centre for Genomics (Sydney, NSW) or the Western Sydney University Next Generation Sequencing Facility (Richmond, NSW).