RNA-Seq of Aspergillus flavus
Fungal tissue was ground using a sterile mortar and pestle pre-chilled with liquid nitrogen. Total RNA was isolated from 100mg of fungal tissue using the Qiagen RNeasy(R) Plant Mini Kit following the manufacturers protocol. Mortar and pestle homogenization followed by RNeasy extraction is effective for isolation of total RNA from filamentous fungi like Aspergillus (Bernaldez et al. 2017). We added 5% -mercaptoethanol to Buffer RLC to reduce RNAse activity.  Total RNA was stored at -80 C until quantity assessment with a Qubit Fluorometer and quality assessment with the Agilent Bioanalyzer. All samples were enriched using a poly-a RNA enrichment and each experiment was pooled separately for sequencing at the University of Arizona Genetics Core