GSM9229419: CTRL_rep_1, HFF/CTRL, biological replicate 1; Homo sapiens; OTHER
Total RNA was isolated using the Direct-zol Miniprep (Zymo Research) according to the manufacturer's instructions including an extended on-column Dnase digestion for 30 min; The 4sU alkylation reaction was performed by incubating 2 to 5 µg total RNA in 1x RNase-free PBS (pH 8) containing 50% dimethyl sulfoxide (DMSO) and 10 mM iodoacetamide (IAA) at 50oC for 15 min. The reaction was quenched with 100 mM DTT and incubated for 5 min on ice, before the RNA was purified using the RNeasy MinElute Cleanup Kit (Qiagen). Library construction was performed by Novogene (Cambridge, UK). Including mRNA enrichment.