title: GSM8263929: ATL-3; Homo sapiens; RNA-Seq
library_construction_protocol: Purified CD4+ T cells from frozen PBMCs of healthy donors and acute ATL patients were stimulated briefly with 2 mg/ml plate-bound anti-CD3 and 1 mg/ml soluble anti-CD28 in the presence of 500 IU/ml recombinant human IL-2 for 4 hours at 37°C. Cells were harvested and loaded into separate channels of a Single Cell Chip G with reverse transcriptase reagent mixture and 5'gel beads according to the manufacturer's protocol (10X Genomics; Pleasanton, CA) Chips were next loaded into the 10X Genomics Chromium Controller for single-cell partitioning, immediately followed by emulsion recovery from the chip and incubated in a MJ Research PTC-200 Thermal Cycler for the reverse transcription reaction, cDNA isolation and library preparation were completed per the manufacturer's protocol. Isolated cDNA was amplified (11 cycles) and was used for the preparation of a gene expression library with the Chromium Next GEM Single Cell 3' Reagent kits v3.1 (10X Genomics). The quality of the cDNA and library was evaluated using the D1000 high-sensitivity kits on a 2200 TapeStation system (Agilent; Santa Clara, CA)