title: GSM8428019: RNA-seq human CD8+ T  donor2-IL2 -4h; Homo sapiens; RNA-Seq
library_construction_protocol: For RNA-Seq experiments, 5´105 purified human CD8+ T cells were cultured in complete RPMI 1640 medium and stimulated for 4 or 24 hr with 1 µM IL-2 or 0.5 µM (equivalent stoichiometric quantity) Control IC or F10 IC. Total RNA was isolated using Direct-zol RNA Microprep Kit (R2061, ZYMO Research), and RNA-Seq libraries were prepared using 120 ng of total RNA and Kapa mRNA HyperPrep Kit (Kapa Biosystems). Each library was indexed using NEXTflex DNA Barcodes-24 (BIOO Scientific) and amplified by PCR. Amplified libraries between 250-400 bp were recovered using 2% E-Gel pre-cast gels (ThermoFisher), purified with Zymoclean Gel DNA Recovery Kit (Zymo Research), and quantified by Qubit 3 Fluorometer (ThermoFisher). After quantification by Qubit (Invitrogen), barcoded samples were mixed and sequenced on an Illumina NovaSeq system.