title: GSM9239960: U2OS cytosolic extract, input, biol rep 3; Homo sapiens; RNA-Seq
library_construction_protocol: After treatment, samples were centrifuged at 14,000 g for 10 minutes at 4°C. Pellets were washed twice with 20 µl ice-cold PBS and resuspended in RNase-free water. Input samples containing U2OS cytosolic extracts were not pelleted. RNA was extracted using TRIzol reagent (Thermo Fisher Scientific) according to the manufacturer's instructions. RNA libraries for RNA-seq were prepared by using poly(A) selection for mRNA enrichment. Following fragmentation, cDNA was synthesized using random hexamer priming. Second-strand synthesis, 5′ end repair, dA-tailing, adaptor ligation, and PCR amplification were performed.