title: GSM8652669: HC_06; Homo sapiens; RNA-Seq
library_construction_protocol: CD4+ T cells were isolated from peripheral blood samples using the CD4+ T Cell Isolation Kit (Miltenyi Biotec). Total RNA was extracted using the miRNeasy Mini Kit (Qiagen) following the manufacturer's protocol. RNA concentration and integrity were assessed using a Bioanalyzer (Agilent Technologies), and samples with a RNA integrity number (RIN) >8 were used. cDNA synthesis followed by transcriptome library preparation were conducted by NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB), where dUTP were incorporated in the process of the 2nd strand cDNA synthesis, instead of dTTP, that block PCR amplification against the 2nd strand templates. This enables to maintain the strandness of RNA transcripts. A 13-cycle PCR amplification was performed to increase library yield and also to incorporate sample barcodes into the library fragments. Resulting transcriptome libraries were quantified by Qubit DNA HS Assay (Thermofisher) and their fragment size distribution was estimated by TapeStation D1000 ScreenTape (Agilent).