title: GSM9103215: wt-expC-2; Schizosaccharomyces pombe; RNA-Seq
library_construction_protocol: Total RNA was extracted from yeast cells using a standard hot acid phenol protocol, followed by phenol-chloroform-isopropanol (PCI) extraction and DNase digestion RNA-seq libraries were prepared with polyA-tail enrichment using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina, followed by using the NEBNext Ultra II library preparation kit for Illumina complemented with custom synthesized barcoded sequencing adapters provided by the Vienna BioCenter Core Facilities Next Generation Sequencing Facility. PCR was performed as a 50 µl reaction using the Q5 polymerase (NEB). Samples were PCR-amplified to the beginning of the logarithmic phase, as determined using a small-scale qPCR reaction run in parallel (typically around 8-10 cycles). Following quality control by Agilent Fragment Analyzer and quantification by RT-qPCR using a Kapa Library Quantification Kit, libraries were sequenced at the Vienna BioCenter Core Facilities Next Generation Sequencing Core Facility on an Illumina sequencer.