GSM8978884: scVDJ-seq of post-treatment peripheral-blood T cells from HNSCC patients; 1B; Homo sapiens; RNA-Seq
Cryopreserved post-treatment PBMC were thawed, and T cells were isolated using EasySep Human T Cell Isolation Kit (Stemcell) per manufacturer recommendations. Cells were washed in 1× PBS, filtered (70 μm) and quantified using acridine orange/propidium iodide (AO/PI) staining on a Cellometer Auto 2000 (Nexcelom). Cells were concentrated to 1000 cells/μL and loaded onto the Chromium Controller (10X Genomics) with a target of 6,000 cells per sample. Cells were mixed with barcoded gelbeads and 5′ GEM Kit v2 reagents (10X Genomics), and single-cell capture was performed. Following reverse transcription, cDNA was amplified, and gene expression libraries were constructed according to the manufacturer's recommendations. Each fragmented DNA library was loaded into a sequencing lane on a NovaSeq 6000 system (Illumina) and was sequenced with pair-end reads of 75 bp.