title: GSM5684311: OH17_vdj; Homo sapiens; RNA-Seq
library_construction_protocol: Fresh whole blood was collected in 2 ml tubes that treated with diamine tetraacetic acid (EDTA). Peripheral Blood Mononuclear Cells (PBMCs) were isolated from whole blood of samples with Ficoll (GE Healthcare Life Sciences) following the manufacture's protocol. Briefly, blood sample was diluted with D-Hanks (NaCl 8 g/L, KCl 0.4 g/L, Na2HPO4 0.16 g/L, KH2HPO4 0.06 g/L, glucose 1 g/L) in a ratio of 1:1, and then add Ficoll and centrifuged at 2000 rpm for 20 min at room temperature. Enriched PBMCs were washed with D-Hanks plus 2% fetal calf serum (FBS), and then centrifuged at 400 g for 10 min, repeat twice. Then cell numbers and viability were assessed. Cell number and viability were checked after surface protein staining (cell viability > 80%). Then droplet-encapsulation single-cell sequencing experiments were performed, and 10,000 living single cells were loaded onto each of the Chromium Controller (10x Genomics). After droplet-encapsulation, single-cell cDNA synthesis, amplification and sequencing libraries were generated using Chromium Single Cell 5' Feature Barcode Library Kit (10x Genomics), Chromium Single Cell 5' Library & Gel Bead Kit (10x Genomics), and Chromium Single Cell V(D)J Enrichment Kit (Human T Cell, 10x Genomics) according to the manufacturer's instructions. The libraries from each loaded channel (up to eight channels) were multiplexed together and sequenced on an Illumina Novaseq 6000.