GSM5328804: PP1860A: WT_replicate1; Schizosaccharomyces pombe; RNA-Seq
Cells were washed once with H2O and quick-frozen by dripping the cell suspension into liquid nitrogen. Cells were lysed in a 6850 Freezer mill (SPEX SamplePrep) using seven cycles (2 min) at a rate of 10 per second with 2 min cooling time between cycles. RNA was extracted four times with 5 ml phenol: chloroform:isoamyl alcohol (25:24:1, equilibrated with 50 mM sodium acetate, pH 5.2) and once with chloroform:isoamyl alcohol (24:1, equilibrated with 50 mM sodium acetate, pH 5.2). RNA was ethanol precipitated and resuspended in H2O. rRNA was depleted using Illumina's Ribo-Zero Magnetic Kit for Yeast as per standard protocol, followed by Illumina's TruSeq Stranded Total RNA LT Sample Prep Kit using the TruSeq Stranded Total RNA Reference Guide