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Target 100s to 1000s of genes to analyze more samples with less sequencing.
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Evercode TCR successfully uncovered nearly five hundred thousand unique beta chain clonotypes across 8 samples in a single experiment.
Pan T cells were isolated from 8 healthy human donors’ PBMCs and fixed using Evercode Cell Fixation Kit v2 to stabilize the cell structure and protect the RNA transcriptome from degradation. Fixed samples were stored at -80C until all samples were ready for further processing in a single experiment with the Evercode TCR Mega kit. Whole transcriptome and TCR specific libraries were sequenced on Illumina Novaseq 6000 S4 and S2 flow cells, respectively. The data was analyzed with Parse Biosciences Analysis Pipeline v1.0.4.
The assay demonstrated a high sensitivity, detecting TCR alpha chains (75% of cells) and beta chains (84% of cells) in primary human T cells. Median transcripts per cell were 1,420 while median genes per cell were 981 at a sequencing depth of 5,000 reads/cell. Clustering of cells using Seurat 4.0 resulted in all the expected major T cell subtypes in a single experiment, as demonstrated in the UMAP below.
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Experimental Summary Report (HTML)
Clonotype Frequency (TSV)
Barcode Report (TSV)
The Adaptive Immune Receptor Repertoire (AIRR) File (TSV)
Whole Transcriptome Results
Digital Gene Expression (DGE) Matrix (12 GB)
All Gene (CSV)
Cell Metadata (CSV)