CLIMUSH: ITS Sequencing of North American Aerial Spore Mycobiome
To identify the aerial spore community, total genomic DNA was extracted using Qiagen DNEasy PowerSoil kits. The ITS1 rDNA subunit was PCR amplified using KAPA HiFi HotStart polymerase (Roche, Indianapolis, Indiana) using the primer set ITS1F_Nextera (TTGGTCATTTAGAGGAAG*TAA) and ITS2_Nextera(GCTGCGTTCTTCATCGA*TGC), both of which contained Illumina adapters (Integrated DNATechnologies, Coralville, Iowa, USA). For the PCRs, 30 cycles were used, with an annealing temperature of 55C. The samples were sequenced at the University of Minnesota Genomics Center on an Illumina NextSeq 2000 machine to generate 2  300 bp paired-end reads.