GSM8490684: HEK293T cells, ERM, Label_Input, 3G, Rep3; Homo sapiens; RNA-Seq
RNAs were extract following the TRIzol Reagent manufacturer's instructions, digested with 1-2 μL DNaseI at 37°C for 30 min, and then incubated with click reagents consisting of 0.1 mM biotin-azide, 2 mM THPTA, 0.5 mM CuSO4 and 5 mM sodium ascorbate. After 10 min of CuAAC reaction at room temperature, RNAs were purified with RNA Clean&Concentrator kit and eluted with pre-warmed nuclease-free water. The RNA integrity was detected by Fragment Analyzer. Approximately 1 μg RNA was kept as pre-enrichment samples, while the remaining was purified with streptavidin beads and the post-enrichment samples were purified by TRIzol Reagent. Total RNAs were used for library construction and libraries were constructed by NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB, E7770) following the manuyfacturer's instructions.