GSM9497150: HEK293T Cells,MGO Treatment,Rep1; Homo sapiens; RNA-Seq
Following treatment, cells were directly lysed in TRIzol (Invitrogen) and total RNA was extracted according to the manufacturer's protocol. Briefly, cells were lysed directly in TRIzol, followed by chloroform phase separation and isopropanol precipitation. RNA was washed with 75% ethanol and resuspended in RNase-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 500 ng of total RNA with RIN values of 10 underwent polyA selection and library preparation using the TruSeq Stranded mRNA LT Kit (Illumina catalog # RS-122-2102) according to instructions provided by the manufacturer with 8 cycles of PCR. Samples were barcoded and run on a NovaSeq 6000 in a PE100 run, using the NovaSeq 6000 S4 Reagent Kit (200 Cycles) (Illumina). An average of 204 million paired reads was generated per sample. Ribosomal reads represented 0.09-0.32% of the total reads generated and the percent of mRNA bases averaged 92%.