title: GSM7093913: dbp2_TTseq_1; Saccharomyces cerevisiae; OTHER
library_construction_protocol: RNA was extracted using a hot phenol protocol. RNAs were sonicated to an average size of 1 kb in a Covaris sonicator. 4-thiouracil-containing RNAs were biotin-labeled using EZ-link biotin (Sigma) at a final concentration of 0.2 mg/mL for 2 hours at room temperature. Treated RNAs were extracted twice with chloroform and ethanol precipitated. Biotinylated RNAs were enriched using the µMACS Streptavidin kit (Miltenyi) as per manufacturer's instructions. Enriched RNAs were ribodepleted using a Ribominus yeast/bacteria ribodepletion kit (Thermofisher) and size selected using an RNA Clean and Concentrator-5 kit (Zymo Research). Using the NEBNext Ultra II Directional RNA Library kit (NEB)