GSM2574347: MCF7_HG→LG; Homo sapiens; RNA-Seq
Total RNA was isolated from cells using TRIzol solution (Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. All RNA samples were quantified by measuring the absorbance at 260 nm and 280 nm (NanoDrop spectrophotometer, Life Technologies, Carlsbad, CA, USA). The integrity of RNA samples was further analyzed by using the digital electrophoresis system Experion with the “RNA StdSens Kit” (Biorad, Hercules, CA, USA), following the manufacturer's instructions. The run and result analysis were performed by the Experion software. RNA samples with a RNA Quality Indicator (RQI) value ≥ 9 were considered good for the further analysis. Paired-end cDNA libraries were prepared for sequencing on the Illumina Hi-Seq 2000 platform using “TruSeq RNA Sample Preparation kit” (Illumina) according to manufacturer's protocol42. The yield and the size distribution of purified cDNA libraries were assessed using “Qubit dsDNA HS (High Sensitivity) Assay Kit” on Qubit Fluorometer (Life Technologies, Carlsbad, CA, USA) and “Experion DNA 1K Analysis Kit” (Biorad, Hercules, CA, USA) on the Experion system (Biorad, Hercules, CA, USA), respectively. Peak median values were determined at approximately 200-260 bp. Sample concentration and peak median values were used to calculate molar concentration of cDNA libraries prior to sequencing on the Illumina HiSeq 2000