GSM9048060: Untreated sample 1; Cryptococcus neoformans H99; RNA-Seq
Exponential C. neoformans H99 cells from YPD culture were pelleted, washed three times with saline, and adjusted to a final density of ~1 x106 CFU/mL in RPMI medium. Individual aliquots were treated with SQ109 (2 μg/mL) or DMSO (control) for 3 hrs at 35 °C under agitation. Treated yeast cells were collected, and RNA extraction was performed using RiboPureTM-Yeast Kit (Ambion, AM1926, MA, USA) following manufacture guidelines [61, 63]. The quality of RNA samples was validated using NanoDrop One spectrophotometer (Thermo Fisher Scientific, DE, USA). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dTTP for non strand specific library or dUTP for strand specific library.