title: GSM7777662: Peripheral blood NK cells, hashtagged, Donor1; Homo sapiens; RNA-Seq
library_construction_protocol: PBMCs were isolated from peripheral blood via Ficoll density separation, and cryopreserved until use. Thawed PBMCs were labeled for FACS with anti-CD3, CD56, NKG2A, NKG2A, NKp30, and DAPI, and sorted on a BD Aria into seven populations: CD56bright NKG2A+NKG2C-, CD56bright NKG2A+NKG2C+, CD56dim NKG2A+NKG2C-, CD56dim NKG2A+NKG2C+, CD56dim NKG2A-NKG2C-, CD56dim NKG2A-NKG2C+NKp30hi, CD56dim NKG2A-NKG2C+NKp30-. Sorted populations were labeled with unique hashtag oligos according to manufacturer's protocol (TotalSeq-B, Biolegend), and 3,000 cells of each populations were pooled together for a total of 21,000 cells per sample from each donor. The samples were submitted to the MSKCC Integrated Genomics Operation in a solution of 0.04% BSA in PBS without calcium or magnesium. Single-cell sample preparation, library prep, cDNA synthesis, and sequencing for Single Cell Gene Expression 3' v3 and Feature Barcoding performed by IGO according to the manufacturer's kit instructions.