GSM9385137: RNAseq_DMSO_treated_parental_MSTO_cells_rep2; Homo sapiens; RNA-Seq
RNA was isolated from 5x105 cells by standard, TRIzol-based RNA precipitation method as follows. Cells were resuspended in 1ml TRIzol (Ambion). Chloroform was added (200ul) to this lysate and extensively vortexed to achieve a homogenous mixture; then, it was incubated for 3-minutes at room temperature before centrifugation at 14,000g (relative centrifugal force) at 4C for 15-minutes. Aqueous layer was collected from the top and transferred into a new tube (~550ul), and the RNA was precipitated with equal volume of 2-propanol for 20-minutes at room temperature. RNA precipitates were centrifuged at 16,000g for 15-minutes at 4C and supernatant was carefully discarded without disturbing the RNA pellet. RNA pellet was washed with 1ml 75% EtOH and after the wash, dissolved in 30ul nuclease-free water. RNA concentration was determined by nanodrop. Approximately, 500 ng of RNA was used for library synthesis using the TrueSeq RNA Sample Preparation kit v2 (Illumina). Size of the libraries was confirmed using 2200 TapeStation, and the concentration was determined by a qPCR-based method using the Library Quantification Kit (KAPA).