title: RNA-Seq of Cryptococcus gattii VGII ZRG1 null mutant cultured in YNB with zinc chelator TPEN for 2h
design: Cryptococcal cells were pre-grown in YPD medium (30C, overnight) on a rotary platform. YNB and YNB supplemented with 10 M TPEN were inoculated with 1  10e7 cells/mL and incubated for 2 h at 30C on a rotatory platform (200 rpm). Cells were collected by centrifugation  and RNA was isolated using TRIzol (Invitrogen) after cellular lysis by glass beads. mRNA was purified from total RNA samples using a Dynabeads mRNA Purification Kit (Thermo Fisher). Purified samples were used for library construction using a TruSeq Stranded mRNA LT Sample Prep Kit (Illumina). The libraries were sequenced using NextSeq 500/550 Mid-Output Kit v2.5 (Illumina) in a NextSeq 500 Sequencing System (Illumina). Raw data were processed using the Illumina BaseSpace Sequence Hub for deindexing and fastq file recovery.