title: GSM8114604: Pooled CARs 10X Lane 1 Feature Barcode Library; Homo sapiens; RNA-Seq
library_construction_protocol: Human primary CD3+ T cells from two donors expressing two novel CARs, CAR 6 and CAR 9, and one control CAR, 13BBz, were subject to antigen stimulation with mitomycin treated IL13Ra2+ U87 cells four times in vitro every 4 days. Residual tumor cells were removed by sorting for bicistronically expressed EGFP Cells were stained with TotalSeq-C hashing prior to sorting and pooled before encapsulation in two channels of the Chromium Single Cell 3′ v3.1 platform (10X Genomics). Gene expression (GEX) libraries were constructed based on manufacturer' instructions, while hashing antibody libraries were constructed as reported previously. The resulting libraries were pooled at 12.5% antibody to 87.5% GEX before sequencing on an Illumina NextSeq500 to a depth of 34,380 reads per cell.