GSM7949326: healthy individual 2; Homo sapiens; RNA-Seq
To isolate human circulating CD8+ T cells, we collected 10 mL peripheral blood from each subject. Then, the EasySep™ Direct Human CD8+ T Cell Isolation Kit (STEMCELL Technologies SARL), in combination with EasyEights Magnets (STEMCELL Technologies SARL), was used to isolate CD8+ T cells from the blood of patients with SJS/TEN and healthy controls, resulting in a CD8+ T cell suspension with a purity of > 90%. Cells were loaded into a 10x Genomics microfluidics chip and encapsulated with barcoded oligo-dT-containing gel beads using the 10x Genomics Chromium controller according to the manufacturer's instructions. Single-cell libraries were then constructed according to the manufacturer's instructions. Single-cell suspensions were loaded to 10x Chromium to capture at least 2000 single cells according to the manufacturer's instructions of 10X Genomics Chromium Single-Cell 3'kit (V3). The following cDNA amplification and library construction steps were performed according to the standard protocol. Libraries were sequenced on an llumina NovaSeq 6000 sequencing system (paired-end multiplexing run, 150bp) at aminimum depth of 20,000 reads per cell.