title: GSM6285174: Spt20-AID, +DMSO, rep-2; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: nsRNA-seq was done as prevously described in Saleh et al, 2021 [PMID: 34383744]. Briefly, Cells were labelled with 4tU for 6 minutes. Total RNA was extracted using the Masterpure Yeast RNA extraction kit (Lucigen MPY03100) as per the manufacturer's protocol. Total RNA was biotinylated using MTSEA-biotin for 30 mins in the dark at room temperature. Then, the biotinylated RNA was purified using Dynabeads MyOne Streptavidin C1 (ThermoFisher) and then purified using Qiagen RNeasy MinElute Cleanup Kit (#74204). rRNA was depleted from the purified nascent RNA fraction using Terminator™ 5′-Phosphate-Dependent Exonuclease (Lucigen TER51020) as per the manufacturer's protocol. rRNA-depleted nascent RNA was purified and concentrated using RNAClean XP clean beads (1.8:1 beads:sample ratio). TruSeq Stranded Total RNA kit for Illumina