GSM9266907: Cells from PBMC from patient #492; Homo sapiens; RNA-Seq
Samples were cryopreserved in R10 medium. Upon thawing, cells were washed, resuspended, and counted, and viability was assessed. CAR positivity was evaluated by flow cytometry. 10x Genomics Chromium GEM-X Single Cell 3' v4 Gene Expression assay following the manufacturer's protocol (10x Genomics, Pleasanton, CA). Briefly, single-cell suspensions derived from PBMC and CSF were evaluated for viability and concentration prior to loading onto the Chromium X instrument for Gel Beads-in-Emulsion (GEM) generation, enabling the barcoding of individual cells and their transcripts. Reverse transcription and cDNA amplification were carried out within the GEMs, followed by library construction, using the Chromium GEM-X Single Cell 3' Kit (PN-1000691), Chromium GEM-X Single Cell 3' Chip Kit (PN-1000690), and the Dual Index Kit TT Set A (PN-1000215). Final libraries were quantified, pooled, and sequenced on an Illumina NovaSeq X system using the 10B Reagent Kit (100 cycles) to a depth of 100K reads per cell.