title: GSM5369341: H113N_Rep4; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: RNA was extracted from whole yeast cells using hot acid phenol, followed by a further phenol-cholorform extraction and ethanol precipitation. Prior to preparing RNA-seq libraries for Illumina NovaSeq sequencing, contaminating DNA was digested using Turbo DNase (Thermo Fisher). RNA quality was then visually assessed by running on a 1% agarose gel to confirm that minimal degradation had occurred. RNA-sequencing libraries were prepared with the KAPA mRNA Hyper Prep kit (KAPA Biosystems), according to the manufacturer's protocols. RNA-seq libraries were assessed for correct fragment size and the presence of adapter dimers buy running on a 1% agarose gel. Average library sizes of ~270 bp were generated. Libraries were pooled for multiplexed sequencing and further purified using KAPA Pure beads. Total DNA concentration was adjusted to 10 nM for Illumina sequencing.