GSM9378674: HA-SL CD22-CAR transduced, post-thaw, 24hr rest, tr2; Homo sapiens; RNA-Seq
At the end of the assay, dead cells were depleted using a Dead Cell Removal Kit (Miltenyi Biotec, Cat #130-090-101) to achieve greater than 90% viability, and cells were resuspended at 1.25e6/mL. Following manufacturer recommendations, emulsion droplets were constructed using 10X Genomics Chromium X controller with the 3' scRNA-seq On-Chip Multiplexing (OCM) workflow to target 5,000 single cells per condition. Gene expression library preparation was performed using the 10X Genomics Chromium Single Cell OCM 3′ Reagent Kit. Quality control on the generated cDNA was performed using an Agilent 2100 Bioanalyzer using a High Sensitivity DNA Kit (Agilent Technologies). One quarter of the cDNA was used to build the gene expression libraries, and indexed using the 10X, Set TT A index plate. Gene Expression libraries were pooled based off the projected number of cells captured and sequenced using a NextSeq 2000 P4 100c flow cell, aiming for 20,000 reads per sample. Two experimental replicates were performed.