RNAseq of C. noformans not treated
Total RNA was obtained from three biological replicates of C. neoformans cells (H99strain) incubated with minimal medium or Gal-3 (0.1 _g/ml in minimal medium) for24 h at 37¡C. The cells were harvested by centrifugation, frozen in liquid nitrogen,and lysed with the use of mortar and pestle for total RNA isolation. Total RNA wasobtained using Trizol (Invitrogen), treated with RNase Ð free DNA I (Fermentas), andcleaned up using an RNAeasy Kit (Qiagen Biotecnologia Brasil Ltda, Sao Paulo, SP,Brazil) following the manufacturerÕs instructions. The RNA was quantified using aQubit 3.0 Fluorometer (Life Technologies), and analyzed using an Agilent 2100Bioanalyzer system to assess the RNA integrity, which ranged from 9.5 to 9.9(integrity number). The barcoded libraries were prepared and sequenced in the Centerfor Medical Genomics of the Ribeirao Preto Medical School (Ribeirao Preto, Brazil)using the Illumina Truseq mRNA Sample Preparation kit according to manufacturerrecommendations. Subsequent clustering and sequencing were performed withthe Illumina NextSeq 500 system.
SUB3155542
H99C2