GSM1754995: LP-1_DMSO_4h_RNA-seq_rep1; Homo sapiens; RNA-Seq
Cells were counted using a Countess Automated Cell Counter (Life Technologies).  Equal numbers of cells for each sample were used for total RNA isolation: Cells were harvested, lysed directly in 1 mL Trizol Reagent (Life Technologies) and incubated for 5 minutes at room temperature.  200 µL chloroform was added to each sample, and samples were shaken for 15 seconds, followed by a 3 minute incubation at room temperature. Samples were spun at 15,000 x g for 15 minutes at 4 ˚C.  The aqueous layer was removed and RNA was purified using an RNeasy Mini Kit (QIAGEN) with on-column DNase digestion. Equal amounts of ERCC RNA spike-in mix 1 (Life Technologies) were added to each sample, to give the same amount of synthetic spike-in RNA per cell equivalent across samples 500 ng of total RNA were used for library construction using the TruSeq RNA Sample Preparation Kit v2 (Illumina)