GSM8489177: SK-N-SH Over Expression Empty Vector, rep1; Homo sapiens; RNA-Seq
Cells were detached and dissociated using accutase and washed 1x with DPBS before RNA isolation using the RNeasy Mini Kit (Qiagen) according to manufacturer's instructions, including homogenization with QIAshredder columns (Qiagen) and on-column DNAse digest with RNase-free DNase set (Qiagen). The amount of total RNA was quantified using the Qubit 2.0 Fluorometric Quantitation system (Thermo Fisher Scientific) and the RNA integrity number (RIN) was determined using the 2100 Bioanalyzer instrument (Agilent). RNA-seq libraries were prepared with the NEBNext® Ultra™ II Directional RNA sample preparation kit (New England Biolabs) according to manufacturer's instructions. NGS library concentrations were quantified with the Qubit 2.0 Fluorometric Quantitation system (Life Technologies) and the size distribution was assessed using the 2100 Bioanalyzer instrument (Agilent).