title: GSM2805400: WT1-2_mRNA_caffeine; Schizosaccharomyces pombe; RNA-Seq
library_construction_protocol: Nascent RNA was prepared using a nascent RNA isolation protocol for yeast (Carrillo Oesterreich et al. 2010) and Phenol:Chloroform:IAA, 25:24:1, pH 6.6 extraction. PolyA- RNA was obtained using oligo-dT coated magnetic beads binding to polyA+ RNA (Dynabeads mRNA DIRECT Micro Purification Kit, Life technologies). rRNA was removed using the Ribo-Zero Gold rRNA Removal Kit (Yeast, Epicenter/Illumina) from ~5 μg RNA. For cytoplasmic mRNA analysis, total RNA was extracted from the cytoplasmic fraction during the nascent RNA isolation protocol and enriched for polyA+ RNA using oligo-dT coated magnetic beads binding to polyA+ RNA (Dynabeads mRNA DIRECT Micro Purification Kit, Life technologies). Random hexamer primed libraries were prepared with standard Illumina library protocols. Single-end sequencing with 76 bp read length was done. Samples were sequenced in biological triplicates for mock- and caffeine-treatment and in duplicate for wild-type strain SP286.
attributes.tag: GEO Accession
attributes.value: GSM2805400