RNA-Seq of K. phaffii: CBS2612
RNA was isolated from cell pellets from chemostat and retentostat cultivations using the Tri reagent according to the supplier's instructions (Ambion, USA). Cells were disrupted by glass beads using a ribolyser (MP Biomedicals). To remove residual DNA, the RNA samples were treated with the DNA-free-kit (Ambion) according to the manufacturers manual. Subsequently, RNA quality, purity and concentration were analyzed by a Nanodrop spectrophotometer and Bioanalyzer (Agilent). Library generation and sequencing were performed at Vienna Biocenter Core Facility NGS Unit (www.vbcf.ac.at). Ribosomal RNA was depleted from total RNA (NEB). The resulting ribo-depleted RNA was then fragmented, and cDNA libraries generated using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB). Libraries were sequenced as paired-end 150 base pair reads on an Illumina NovaSeq 6000 using the SP workflow.