GSM7466950: Slow_Netseq_Rep2; Saccharomyces cerevisiae; OTHER
3'READS: Polyadenylation sites were detected using the 3'READS method (Jin, Y. et al., 2015). RNA was extracted  by hot acid phenol followed by Qiagen RNeasy purification. 3' READS libraries were prepared as previously described with 25µg total input RNA (Jin, Y. et al., 2015). NET-seq: NET-seq samples were prepared as described previously (Churchman and Weissman,2012). Yeast grown to mid-log phase were isolated by filtration, and flash-frozen in liquid nitrogen. Frozen cell pellets were cryogenically lysed by six three-minute cycles of grinding in a cryomill . Pol II-RNA complexes were affinity purified using anti-FLAG agarose slurry.  . NET-seq:  Isolation of nascent RNA and library construction for NET-seq samples was performed as described in Couvillion et al., (2022). Library concentration and size distribution was determined using a 2100 Agilent Bioanalyzer and qPCR quantitation with library-specific primer (TCCGACGATCATTGATGGTGCCTACAG).