title: GSM3182554: Cryptococcus neoformans H99 treated with eltrombopag; Cryptococcus neoformans var. grubii H99; RNA-Seq
library_construction_protocol: After this treatment, cultures were centrifuged and total RNA was extracted using Trizol reagent and treated with DNase I (Ambion, Thermo Fisher Scientific) to degrade contaminating DNA. mRNAs were enriched with oligo (dT) magnetic beads. Fragmentation buffer allowed the mRNA to be shortened into ~200-base fragments. Then, the first strand of cDNA was synthesized using random hexamer, buffer, dNTPs, and RNase H, and the second strand of cDNA was synthesized by DNA polymerase I. The double-stranded cDNAs were purified with magnetic beads. End preparation and 3' end single nucleotide adenine addition was performed, and sequencing adaptors were ligated to the fragments, which were enriched by PCR amplification. During the quality control step, the Agilent 2100 Bioanaylzer and ABI StepOnePlus RealTime PCR system were used to qualify and quantify the sample library. The library products were sequenced via Illumina NextSeq 500.