GSM8325605: TTseq_Aux4h_NVP2_30m_ELOA-WT_NELFC-AID_Rep1; Homo sapiens; OTHER
TT-seq library was prepared as described previously (Schwalb et al., 2016; Rosencrance wt al., 2020) with these modifications:  after treatment cells were immediately lysed in TRIzol and the lysates were stored at –80°C. ~150 µg total RNA supplemented with 4sU-labeled fly S2 spike-in RNA was fragmented at 94°C for 2 min using NEBNext Magnesium RNA Fragmentation Module. The labeled RNA was biotinylated using 33 µg/ml MTSEA-biotin-XX in the presence of 20% DMF for 1 h at r.t. The biotinylated RNA was purified using Dynabeads MyOne Streptavidin C1 and eluted in 100 mM DTT. rRNA was depleted using NEBNext rRNA Depletion Kit v2. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit