title: GSM8070458: ctr9D, 4tU, Rep2, Paf1CD; Schizosaccharomyces pombe; Saccharomyces cerevisiae; OTHER
library_construction_protocol: 4tU-labeled S.cerevisiae pellets were spiked-in with labeled S. pombe at a 14:1 OD600 ratio and total RNA extracted with a hot acid-phenol extraction. 4tU-RNA labled was biotinylated with MTSEA-XX biotin and pulled down using Dynabeads MyOne Streptavidin C1 beads. RNA was purified using the RNEasy kit and quantified by Qubit. 4tU-seq libraires were prepared using the TECAN Ovation SoLo RNA-seq library preparation kit (TECAN, Redwood City, CA; 0516–32) targeting S. cerevisiae rRNAs for depletion using AnyDeplete technology. 2-10 ng of RNA was used per library build and fragment distributions assesed by tapestation