GSM4476445: Th17_highsalt_2; Homo sapiens; RNA-Seq
Total RNA was extracted from cells lysed in TRI reagent (Sigma-Aldrich) according to the manufacturer's protocol. RNA was quantified using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific) and its quality was verified by an Agilent 2100 Bioanalyzer (Agilent) according to the manufacturer's guidelines. Library preparation for RNA-seq was performed using the TruSeq® Stranded Total RNA Sample Preparation Guide (Illumina) starting with 450 ng RNA as the input for each sample. Only 11 cycles were used for PCR amplification to minimize PCR bias. The barcoded libraries were sequenced on a HiSeq 2500 (Illumina) with paired-end, 100-bp reads. Approximately 6 Gb of sequencing reads were produced on average per sample.