TCR Sequencing of 1 Million Mouse T Cells in a Single Experiment

Key Takeaways

• 1 million T cells profiled in a single experiment
• Joint measurements of gene expression and paired TCR alpha and beta chains in single cells
• Characterization of TCR diversity across the mouse lifespan

Fresh spleens of mice from 1 month through 27 months were dissociated into single cell suspensions and T cells were isolated using magnetic bead-based negative selection. T cells were fixed independently using an Evercode Cell Fixation to stabilize the cell structure and protect the RNA transcriptome from degradation and fixed cells were stored at -80C prior to barcoding and library prep.

1 month, 2.5, 6, 9, 12, 17, and 27 months old mice spleens were processed with Evercode TCR Mega. Sequenced on the Illumina NovaSeq X at ~10,000 mean reads per cell. Sequencing data were processed with the Parse Biosciences pipeline v1.3.0, clustered with Seurat v5, annotated, and visualized as a UMAP. Whole transcriptome-based clustering accurately identified low abundance non-T cell types and major T cell populations as shownin the UMAP below.

Next Steps

• Review a more comprehensive description of Evercode TCR in the Evercode TCR Technical Brochure.
• Visit the Evercode TCR product page for Evercode TCR product highlights.