GSM6340855: shLUC_rep1; Homo sapiens; RNA-Seq
Approximately 50 mg of fresh-frozen primary tumor tissue was crushed in liquid nitrogen in presence of QIAzol Lysis Reagent (Qiagen, Hilden, Germany). The total RNA was isolated using miRNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. All RNA samples were treated by DNase I during the isolation procedure. The RNA yield was determined by UV absorbance using a NanoDrop 1000 spectrophotometer (Peqlab, Erlangen, Germany). The RNA quality was assessed by analyzing the ribosomal RNA integrity number (RIN) on an Agilent 2100 Bioanalyzer using the RNA 6000 Nano kit (Agilent Technologies, Palo Alto, CA, USA). The RIN values of the isolated RNA samples ranged from 6.7 to 9.6. Libraries for mRNA sequencing were prepared from total RNA samples using the Illumina Stranded mRNA Library Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions.