GSM9017309: WT 293T, rep2; Homo sapiens; RNA-Seq
Frozen cells were lysed in TRIzol Reagent (ThermoFisher catalog # 15596018) and phase separation was induced with chloroform. RNA was precipitated with isopropanol and linear acrylamide and washed with 75% ethanol. The samples were resuspended in RNase-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 500ng of total RNA with RIN values of 9.7-10 underwent polyA selection and TruSeq library preparation according to instructions provided by Illumina (TruSeq Stranded mRNA LT Kit, catalog # RS-122-2102), with 8 cycles of PCR. Samples were barcoded and run on a HiSeq 4000 in a PE50/50 run, using the HiSeq 3000/4000 SBS Kit (Illumina).  An average of 47 million paired reads was generated per sample. Ribosomal reads represented 0-3.8% of the total reads generated and the percent of mRNA bases averaged 72%.