title: 454 GS FLX+ sequencing
design: Fungal intergenic transcribed spacer region of  rDNA gene, ITS33
library_construction_protocol: triplicate PCR reaction (ITS1F/ITS4) final volume 50ul containing 1xGoTaqGreen Mastermix (Promega), 25pmol of each primer, 10ng DNA; initial activation 95C for 5 min, 10 cycles , QIAquick Gel Extraction Kit (Qiagen), Quant-iT PicoGreen dsDNA Assay Kit, equimolar pooling of samples, sequencing from gene end, trimming to 350bp containing ITS2 region, normalization, cd-hit clustering, GAST taxonomic assignment, uchime chimera check