GSM5584161: Nkcells_sample7-17 [S11 S18]; Homo sapiens; RNA-Seq
Peripheral Blood Mononuclear Cells (PBMCs) were isolated with density gradient seperation. Cells were subsequently incubated with barcoded (hashtaging) and fluorochrome-conjugated antibodies and NK cells sorted by FACS as live CD45+ CD56+ cells. These cells were then applied to the 10X Genomics platform using the Single Cell 5' Library & Gel Bead Kit (10x Genomics) following the manufacturer's instructions. The purified cDNA was amplified using 12 or 13 cycles of PCR and then used for 5' gene expression (GEX) and cell-surface hashtaging library preparation. Through fragmentation, adapter ligation and index PCR the final libraries were obtained. The quality of single cell 5' GEX was assessed by Qubit quantification and Bioanalyzer fragment analysis (HS DNA Kit, Agilent). The sequencing was performed on a NextSeq500 device (Illumina) using High Output v2 Kits (150 cycles) with the recommended sequencing conditions for 5' GEX and cell-surface hasthgaing libraries (read1: 26nt, read2: 98nt, index1: 8nt, index2: n.a.).