GSM8987451: H99YPD3; Cryptococcus neoformans H99; RNA-Seq
Total RNA was extracted from freshly collected fungal cells using TRIzol reagent (Invitrogen) following the manufacturer's protocol. RNA quantity and integrity were assessed using an Agilent 2100 Bioanalyzer, and only samples with a RNA Integrity Number (RIN) >7.0 were used for library construction. Poly(A)+ mRNA was enriched using oligo(dT) magnetic beads and then fragmented using a fragmentation buffer. cDNA was synthesized using random hexamer primers and reverse transcriptase. The resulting cDNA was purified, end-repaired, A-tailed, ligated with Illumina sequencing adapters, size-selected, and PCR-amplified to construct the final sequencing libraries. Libraries were sequenced on the Illumina NovaSeq 6000 platform to produce 150 bp paired-end reads according to the standard protocols by Majorbio Bio-Pharm Technology Co. Ltd., generating an average of ~50 million high-quality reads per sample.