GSM5931532: PBS rep1; Mus musculus; RNA-Seq
After exposure, the cells were lysed in 500 μL of cold cell lysis buffer containing 1% LiDS, 100 mM Tris-HCl pH 7.5, 500 mM LiCl, 10 mM EDTA, and 5 mM DTT (Shichino et al., 2019). The cell lysates were stored at -80 °C until analyses. Transcriptome libraries were prepared on cell lysate resolved in lysis buffer. PolyA RNAs were isolated using Dynabeads M-270 Streptavidin (Thermo Fisher Scientific) conjugated with biotin-3' WTA-EcoP-dT25 according to the previous report with some modifications (GSE110711). 100 ng of the whole-transcriptome library was subjected to fragmentation/end-repair/A-tailing using NEBNext Ultra II FS DNA Library Prep Kit for Illumina (New England Biolabs) with some modifications. The thermal cycling was performed as following condition: for 20 min at 32ºC, 30 min at 65ºC, and hold at 4ºC. Then, 1.25 μL of 1.5 μ