GSM8412930: Human induced Treg cells, transduced with empty vector (EV), donor 2; Homo sapiens; RNA-Seq
Total RNA was purified using the miRNeasy Micro Kit (QIAGEN). cDNA was reversely transcribed from total RNA with oligo-dT primer, random hexamers with SuperScript™ II Reverse Transcriptase (Invitrogen) and then amplified with IS PCR primers and Kapa HiFi Hotstart Ready Mix (Roche). Then the cDNA library was fragmented and tagged with sequencing adapters using Nextera XT DNA Library Preparation Kit (Illumina), and finally ~500 bp amplicons were got. Quality of library was assessed using the High Sensitivity RNA ScreenTape Kit (Agilent Life Sciences). DNA was quantified using Quant-iT PicoGreen dsDNA Assay Kits (Invitrogen) and purified using AMPure XP beads (Beckman Coulter). Libraries were subjected to 2 × 50-bp paired-end sequencing on an Illumina NovaSeq6000 (Illumina), generating at least 8 million uniquely mapped reads for each sample.