title: GSM5107257: SPAC20G4.07c_G12_rep3; Schizosaccharomyces pombe; RNA-Seq
library_construction_protocol: Total RNA extraction was performed according to [A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae, Mark E.Schmitt, NAR, 1990] and the RNA pellet was resuspended in 87ul of RNase free water. DNA was digested using 3ul of DNase (Qiagen) and 10ul RDD DNase buffer at 37ºC for 30 minutes. RNA cleanup was performed with the Purelink RNA Mini kit (Thermo Scientific) according to the manufacturer's instructions. RNA integrity was measured on a Bioanalyzer 2100 system (Agilent). NGS library prep was performed with Lexogen 3´mRNA-Seq Library Prep Kit FWD HT following Lexogens standard protocol (015UG110V0110). Libraries were prepared with a starting amount of 338ng and amplified in 13 PCR cycles. Libraries were profiled using a High Sensitivity DNA chip on a 2100 Bioanalyzer (Agilent) and quantified using the Qubit dsDNA HS Assay Kit in a Qubit 2.0 Fluorometer (Life Technologies). 2x192 libraries were pooled together in equimolar ratio and sequenced in single-end mode on 2 NextSeq 500 High-output flowcells, for 1x75 cycles plus 2x8 cycles for the dual index read.