GSM8125716: EUseq, aux, rep2; Homo sapiens; OTHER
Total RNA from HCT116 cells and drosophila S2 cells was isolated with Trizol (Invitrogen) and Nucleospin RNA II (Macherey-Nagel) following the manufacturer's instructions. rRNA was depleted from total RNA from HCT116 cells with NEBNext rRNA Depletion Kit (NEB). mRNA was purified from total RNA from S2 cells with Dynabeads mRNA Purification Kit (Invitrogen). 200 μg of rRNA-depleted RNA of HCT116 cells and 20 μg of mRNA of S2 cells were mixed, and EU-labelled RNA was isolated with Click-iT Nascent RNA Capture Kit (Thermo Fisher). Single-strand cDNA was synthesized from the isolated RNA by SuperScript VILO cDNA synthesis kit (Thermo Fisher). Second-strand cDNA synthesis and the subsequent steps to create library were conducted with NEBNext Ultra II Directional RNA Library Prep Kit for Illumina and NEBNext Multiplex Oligos for Illumina (New England BioLabs).