GSM7509631: TT-TL_HEK293T_TSA_2; Homo sapiens; RNA-Seq
For TT-TL-seq, RNA was harvested with Trizol reagent, followed by chloroform extraction and isopropanol precipitation. Genomic DNA was depleted using Turbo DNAse, followed by cleanup with RNAclean beads. 50 μg of total RNA was subjected to MTS chemistry, followed by biotinylation and streptavidin enrichment as previously described (Duffy et al., 2015). Nascent enriched RNA for TT-TimeLapse-seq RNA was then treated with oxidative-nucleophilic-aromatic-substitution chemistry using mCPBA as the oxidant as previously described (Schofield et al., Nature Methods 2018). TT-TL-seq libraries were generated following standard protocols using the SMARTer Stranded Total RNA-Seq Kit-Pico Input Mammalian, V2 (Takara Bio USA, cat. 634413).