title: GSM3573472: n_RNA-WT_dmso-rep1; Saccharomyces cerevisiae BY4741; RNA-Seq
library_construction_protocol: Total RNA was extracted using a standard hot acidic phenol approach. Prior to lysis, S. pombe cells were added in a 1:6 pombe-to-cerevisiae cellular ratio. Pombe cells were also labeled with 4-thiouracil. RNA was treated with DNase I (Ambion, #AM1906). For nascent RNA extraction, 100 ug of total RNA was biotinylated with 10 ug of MTSEA-biotin XX (Biotium, #90066-10mg), and the labeled RNA was purified using Dynabeads MyOne Streptavidin C1 beads (Thermo Fisher Scientific, #65001). Both total and nascent RNA were ribosomal RNA-depleted and prepared for sequencing using the Yeast ScriptSeq Complete Gold Kit (Epicentre, BGY1324). DNA was sequenced on an Illumina HiSeq 2000 generating single-end reads of 100 bp length using standard Illumina protocols.