RNA-seq of S. cerevisiae S288C wild-type and S288C-H2 mutant
RNA-seq of S. cerevisiae S288C wild-type and S288C-H2 mutant
Cultures were incubated at 30 ˚C with shaking (220 rpm). Samples of 1.5 mL from each culture were collected at 2 h, 4 h, 6 h, 8 h, 12 h, 2 4 h, 48 h and 72 h, centrifuged for 5 minutes at 8000 g, and, after discarding the supernatant, pellets were stored at -80 ˚C until further processing.  RNA was extracted from the yeast by a lysis protocol using a combination of sonication at 20 kHZ with a 50% amplitude for 10 seconds + TRIzol® Reagent (Invitrogen, Waltham, MA, USA) following manufacturer's instructions. RNA purification was performede using PureLink RNA Mini Kit™ (Invitrogen) following manufacturer's instructions. RNA concentration and purity was determined using the NanoDrop™ 2000 spectophotometer (Thermo Scientific). RNA integrity was determined by capillary electrophoresis at the TapeStation using Agilent High Sensitivity RNA ScreenTape Assay (Agilent). Illumina Stranded mRNA libraries were prepared using the TruSeq Stranded mRNA kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions.