RNA-seq of Schizosaccharomyces pombe
Wild-type cells (972 h-) of S. pombe and TUT-ase deletion strains were grown in YES at 32C to the early exponential growth phase (OD 0.5). Total RNA was isolated using the hot phenol method. RNA quality was assessed on a Bioanalyzer instrument (Agilent), treated with DNase (Turbo DNA-free, Ambion), and subsequently, 4 g of RNA was treated with a beta version of Ribo-Zero Magnetic Gold Kit Yeast (Epicentre) to deplete rRNAs. RNA-seq libraries were prepared from rRNA-free RNA using a strand-specific library preparation protocol (a customised version of Illumina TrueSeq Small RNA)