title: ITS2 amplicon sequencing of ratiata pine seedling root-associated fungal communities
design: Pinus radiata seedling teated with amino-acid or inorganics soursce of N fertiliser.  lateral roots (~100 mg ) subsampled from 6 month old seedlings for DNA extraction using the ISOLATE II Plant DNA Kit (Meridian Bioscience) according to the manufacturers protocol. The fungal ITS2 region was amplified by a two-step PCR: first with tailed ITS2 primers (Forward: 5-CCTACACGACGCTCTTCCGATCTNNNNGTGARTCATCGAATCTTTG-3; Reverse: 5-GTGTGCTCTTCCGATCTCCTCCGCTTATTGATATGC-3), then with indexed Illumina adapters. Libraries were pooled equimolarly and sequenced on an Illumina MiSeq using the v3 600-cycle kit (2300 bp, paired-end).