For data analysis, UMIs and Raw Reads are used to ensure high precision around each clonotype sequence identified. Even when present at only 0.01%, the Jurkat RNA is readily quantifiably identified. Table 1 shows the number of raw reads and the demultiplexed unique captures (UMIs) per Jurkat TCR-alpha and TCR-beta clonotype. Sensitive to at least 0.01% RNA from Jurkat cells was spiked into RNA extracted from peripheral blood mononuclear cells (PBMCs Precision Medicine) at 10%, 1%, 0.1% and 0.01% and used to make an RNA-seq library. The data analysis included with the purchase of the QIAseq Immune Repertoire T-cell receptor panels includes an online portal that seamlessly integrates with Illumina BaseSpace and provides primary read mapping, UMI demultiplexing and reports on sequencing performance, TCR chain usage, CDR3 peptide sequence and length distributions, together with rarefaction and V/D/J usage heat maps. This figure shows the major clonotype of the Jurkat cell, as well as the diversity of the PBMC background. Comprehensive view of the T-cell immune repertoire The heatmaps allow for easy identification of enriched clonotypes across the sample.
