Environmental DNA sequences collected from soil
The ITS region was amplified using primers ITS9mun (5'-TGTACACACCGCCCGTCG-3') and ITS4ngsUni (5'-CCTSCSCTTANTDATATGC-3') for identification of eukaryotes in general. Both primers were equipped with unique 12-base Golay barcodes for multiplexing. Primer pair ITS9mun and ITS4ngsUni has been proposed for analysis of eukaryotes with approximately species-level resolution. These primers amplify 170 bp of the 18S rRNA gene and the full-length ITS region, which serves as an official barcode for fungi and performs well on many other groups including plants.