GSM8579440: EW8, DMSO, rep2; Homo sapiens; RNA-Seq
Cells were harvested and the suspension was centrifuged at 800x g for 5 minutes in decant culture media. The cell pellet was washed in cold PBS and spun down again at 800xg for 5 minutes. Total RNA was extracted using the Rneasy Micro kit. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.