Although sequencing of bulk T cell samples has helped to further understanding of T-cell receptor (TCR) repertoire diversity, bulk sequencing cannot determine the pairing of specific alpha-beta receptor chains within the population of T cells. Since the unique alpha-beta pairing of a TCR mediates antigen specificity, obtaining pairing information will be crucial for providing insights on antigen recognition. While sequencing of single T cells can determine the pairing of a given cell, it can be prohibitively expensive and involves complicated analyses since most methods require sequencing of a large number of cells.

With the SMARTer Human scTCR a/b Profiling Kit, we solve this problem by performing SMART cDNA synthesis with a set of unique indexed oligos that allow the condensing of 96 samples, which have been sorted manually or via FACS, into twelve pools for sequencing. Additionally, the approach enables the twelve pools to be further multiplexed such that all 96 samples are run in a single flow-cell lane. Much like the SMARTer Human TCR a/b Profiling Kit for bulk samples, this kit combines SMART cDNA synthesis and RACE-based PCR, followed by TCR gene-specific PCR to fully capture and amplify TCR-alpha and TCR-beta variable regions to generate Illumina-ready libraries that provide a highly sensitive approach to sequencing TCRs.

If you are looking for the software to demultiplex your sequencing data generated with this kit, please visit the SMARTer Human scTCR Demultiplexer webpage.