title: GSM8816794: Aph, peripheral blood (PB) stem cell apheresis products collected three years prior to CAR-T infusion, GEX; Homo sapiens; RNA-Seq
library_construction_protocol: Samples were thawed and quality was assessed by acridine orange and propidium iodide (AO/PI) staining using automated cell counting. Samples with initial viability <80% were subjected to dead cell removal using ViahanceTM dead-cell removal kit (BioPAL, Inc., United States). The cells were stained with a custom panel of 70 TotalSeqTM-C antibodies (BioLegend, United States), optimized, and prepared by Singleron Biotechnologies, Singapore. These TotalSeqTM-C antibodies are oligonucleotide-coupled antibodies directed against selected cell surface proteins. The antibodies are conjugated via a linker to oligonucleotides, which consist of a polymerase chain reaction (PCR) handle, an antibody-specific barcode, and a capture sequence. These antibody-bound oligonucleotides can be captured during single-cell library preparation, allowing simultaneous surface protein and gene expression profiling. The single-cell gene expression, surface protein expression, and T-cell receptor (TCR) libraries were constructed using Chromium single-cell 5' reagent kits (v2 chemistry; 10x Genomics, Inc., United States), as per the manufacturer's instructions. Libraries were sequenced on an Illumina NovaSeq X platform using a 10B flow cell and PE150 chemistry.
identifiers.namespace: GEO
identifiers.id: GSM8816794_r1