title: 5EU-treated Hela cells, 2 h, Replicate 2
design: Library preparation for direct RNA sequencing was performed as previously described (Ibrahim et al., 2021) with modifications. Briefly, poly(A) RNAs were purified from 50 g of total RNA using Oligo d(T)25 Magnetic Beads (New England Biolabs). 500 ng of poly(A) mRNA was used for library preparation using SQK-RNA002 sequencing kit (Oxford Nanopore Technologies). The final library was quantified using Qubit dsDNA High Sensitivity assay kit (ThermoFisher) and sequenced on a MinION device using FLO-MIN106 flow cells (Oxford Nanopore Technologies)