Fungal ITS amplicon, Granite Mountains, Mojave National Preserve, Dirt clinging to moss biocrust
DNA was extracted using the QIAGEN DNeasy PowerSoil kit (Qiagen, Germantown, MD, USA) from .25 g of sample. The manufacturers protocol was used with a ten minute extension of bead beating. PCR was performed using ITS1F and ITS2 primers targeting the ITS1 region of fungal ribosomal DNA in accordance with the Earth Microbiome Project (EMP). PCR products were normalized and cleaned using the Invitrogen SequalPrep kit (Invitrogen, Waltham, MA, USA). The resulting pools were quantified using an Agilent 2100 Bioanalyzer and Fragment Analyzer (Agilent Technologies, Santa Clara, CA, USA) and qPCR. The following primer constructs were used for sequencing: Read 1 primer, Read 2 primer, and index primer per the EMP protocol for ITS amplicon sequencing (P. Smith et al. 2018). Libraries were sequenced using the illumina MiSeq platform (Illumina, Inc, San Diego, CA, USA)