GSM7745534: SKOV3 control, repeat 3; Homo sapiens; RNA-Seq
Total RNA was isolated from cancer cells using RNeasy Mini Kit (Qiagen) followed by treatment with DNAse I (NEB). The reaction was purifed with PureLink RNA Mini Kit (ThermoFisher). RNA quality was checked by 2100 bioanalyzer (Agilent, USA). Enrichment of polyadenylated RNA and library preparation was performed with NEBNext Ultra II Directional RNA Library Prep Kit (NEB), according to the manufacturer's protocol. The library underwent a final cleanup using the Agencourt AMPure XP system (Beckman Coulter) after which the libraries' size distribution and quality were assessed using a high sensitivity DNA chip (Agilent Technologies). Libraries were subsequently quantified by Quant-iT DNA Assay Kit, High Sensitivity (ThermoFisher). Samples were sequenced on NextSeq 500 System (Illumina, USA) with NextSeq 500/550 High Output Kit v2.5 (75 Cycles). RNA libraries were prepared for sequencing using standard Illumina protocols