title: GSM8035299: siCTRL C4-2  rep1_12h; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA (two replicates at siCTRL, siFBL at  0h, 6h,12h,24h) was isolated from C4-2 cells RNA  by Trizol (Thermo), and the 4-thiol groups present on S4U-labeled transcripts were alkylated with iodoacteamide. The mRNA degradation kinetics was measured using SLAM-seq Kinetics Kit: Catabolic Kinetics Module (LEXOGEN) by following the manufacturer's protocol. In general, control or FBL-deficient C4-2 cells were cultivated in 1640 medium containing 100 μM 4-thiouridine (S4U) for a continuous 24-hour duration. Subsequently, the S4U-containing medium was exchanged by medium with an excess (10 mM) of unmodified uridine (U), and cell samples were collected at the specific time-points. The reference point for these time-points was set at 0 hour, corresponding to the moment of S4U removal. RNA isolation was performed on all collected samples by Trizol (Thermo), and the 4-thiol groups present on S4U-labeled transcripts were alkylated with iodoacteamide. The libraries were generated from 70-200 ng of input RNA using QuantSeq 3' mRNA-Seq Library Prep Kit (LEXOGEN).