DriverMap™ AIR TCR Profiling Kit (Human RNA), V2

Product Description

The DriverMap™ AIR TCR Profiling Kit (Human RNA) is an easy-to-run, single-day assay that uses multiplex PCR-NGS technology to profile the T-cell receptor (TCR) repertoire.

• Profile all TCR chains (TRA, TRB, TRG and TRD)
• Includes all the enzymes, reagents, and primers to prepare 24- or 96- samples ready for NGS from total RNA
• Optimized RT-PCR workflow. No specialized equipment required
• Run in parallel with the DriverMap AIR TCR (Human DNA) kit with DNA starting material for insights into activated clonotypes
• Data compatible with analysis using MiXCR (MiLabs) immune receptor profiling software and similar analysis packages

How It Works

The DriverMap™ AIR TCR Profiling Kit combines multiplexed RT-PCR amplification with the depth and precision of Next-Generation Sequencing (NGS) to quantitatively measure gene expression and analyze RNA sequences of all functional immune receptor mRNA isoforms for TCR chains (TRA, TRB, TRD, TRG) with the exclusion of non-functional pseudogenes and ORFs (as defined by IMGT database).

The AIR assay allows profiling of the repertoire of antigen-binding CDR3 regions using a set of experimentally validated forward primers (designed for variable FR3 region), and reverse primers (designed for the conserved C region) of TCR mRNAs.

The CDR3 region determines the structure and specificity of the antigen. The majority of repertoire profiling studies are based on the analysis of only the CDR3 region, as this is the hypervariable region and contains crucial information to study immune repertoire diversity.

Key Applications

• Tracking T cell diversity and clonality
• Finding markers of autoimmune diseases
• Studying immune cell migration patterns
  
  

Fig.1. Outline of DriverMap AIR RT-PCR-NGS assay for expression and sequence analysis of immune receptor mRNA regions.

Supporting Data

Fig.2. Number of clonotypes for 7 TCR/BCR chains identified in 50ng of normal PBMC or whole blood RNA (10x10^6 reads per sample, triplicates).

Fig 3: Comparison of TCR clonotypes detected by DriverMap AIR RNA full-length assay vs SMART-based assay. Both assays were run with 50 ng total RNA isolated from PBMCs. Data shows that the DriverMap AIR RNA assay detects approximately 3X more TCR clonotypes than the SMART-based assay.