GSM9111066: Purified CD4+ lymphocytes, sample 84T1; Homo sapiens; RNA-Seq
Isolation of CD4+ cells from peripheral blood (PB) was performed using the untouched human CD4+ T Cell Isolation Kit (Miltenyi Biotech, Germany) following the manufacturer instructions. Purity of samples measured by FACS assessed >90% of CD4+ T cells for each patient analyzed in this study. Direct-zol RNA Miniprep (Zymo Research) obtained high-quality total RNA according to the manufacturer's instructions. An equimolar libraries, measured by Bioanalyzer High Sensitivity DNA 1000 Assay and Qubit® RNA HS Assay Kit, was loaded onto Illumina NextSeq 500 platform according to the manufacturer's recommendations. To obtain a high percentage of sequencing reads, we pooled 8 library samples per run into a High Output Kit v2.5 cartridge (paired-end sequencing with about 50 million clusters per sample).