GSM6753135
Exponentially grown yeast cultures (OD600 = 0,8) were isolated from a 5 ml culture sample (3,000 rpm; 3 min, 4°C), washed with ice-cold AE buffer (50mM sodium acetate pH 5.2, 10mM EDTA pH 8.0), and stored at -80°C. Next, total RNA was extracted using phenol and chloroform. Residual genomic DNA was removed (1h, 37°C) with DNase I (New England Biolabs, Cat # M0303L). All cytoplasmic and mitochondrial rRNA were removed from one µg of purified yeast RNA (see above) with the QIASeq FastSelect rRNA Yeast (Qiagen, Cat # 334215). RNA-Seq libraries were synthesized with NEBNext Ultra II Directional Library Prep Kit for Illumina (NEB, Cat # E7760S) using Unique Dual Index UMI Adaptors RNA Set 1 (NEB #7416).