GSM9589545: HEK 293, WT_52x_1rep; Homo sapiens; OTHER
Total RNA was isolated using TRIzol/chloroform/isopropanol method. 50 ng RNA per sample was used for library construction. Newly synthesized RNA was labeled with 4-thiouridine (4-SU) and co-extracted with total RNA. The RNA was fragmented, and 4-SU labeled RNA was biotinylated and purified using streptavidin pull-down. First-strand cDNA synthesis was performed using random hexamer primers, followed by second-strand cDNA synthesis incorporating dUTP to generate strand-specific libraries.