Illumina MiSeq of environmental soil sample
DNA was extracted from soil using Fast Spin Soil DNA kit.  The fungal ITS sequence was amplified with 2 different Nextera-tagged primers (ITS86F with ITS4R; and ITS3F with ITSOF4R).  Amplicons were pooled, cleaned, and used for second round PCR with Nextera adaptor primers.  The resultant amplicons were used in Illumina MiSeq.