title: GSM6668788: 4tU-Seq Replicate 2; Saccharomyces cerevisiae; OTHER
library_construction_protocol: Hot phenol RNA extraction. 4tU-RNA enrichment was performed similarly as previously described (Duffy and Simon, 2016) with minor modifications. Briefly, polyA RNA was obtained from hot-phenol extracted RNA, fragmented with Magnesium RNA Fragmentation Module (NEB) and biotinylated with activated disulfide methane thiosulfonate (MTS) biotin (Biotium Biotin-XX MTSEA). Alternatively, 4tU-RNA was purified from hot-phenol extracted RNA and then subjected to polyA enrichment and magnesium fragmentation. Fragments of mRNA were ligated to a linker and a cDNA library was prepared as described (Jin et al., 2015).