GSM7928817: FBY2578_2_WT; Schizosaccharomyces pombe; RNA-Seq
Total RNA was extracted by the hot-phenol method, as previously described (Duval et al., 2023). Briefly, yeast cells are first resuspended in 600 µl of TES solution (10 mM Tris-HCl [pH 7.5], 10 mM EDTA [pH8.0] and 0.5% SDS) and 600 µl of phenol-chloroform-Isoamyl alcohol (125:24:1 [pH 4.5]), and then incubated at 65°C for 60 min with vortexing every 10 min. The RNA was extracted once with 550 µl of phenol-chloroform-Isoamyl alcohol (125:24:1 [pH 4.5]) and once with 450 µl of chloroform-isoamyl alcohol (24:1), and precipitated with ethanol and 2 M NaCl. rRNA-depleted RNA-seq libraries were prepared following manufacturer's instructions using the Illumina Truseq Stranded Total RNA-sequencing kit after ribodeletion with the RiboZero Yeast ribodepletion (Epicentre). RNA-seq libraries were sequenced in paired-end (2×100nt) using Illumina technologies (HiSeq 4000) at Genome Québec.