Takara SMARTer Human TCR a/b Profiling Kit v2 \48 Rxns

The SMARTer Human TCR a/b Profiling Kit v2 enables users to analyze T-cell receptor (TCR) repertoires from bulk RNA samples. This kit can generate libraries from total RNA (10 ng to 1 µg from peripheral blood leukocytes, 20 ng to 200 ng of total RNA obtained from whole blood, or 1 ng to 100 ng T cells) or purified T cells (1,000 to 10,000 cells) and can be used to generate data for both TCR-alpha or TCR-beta subunits. Unique molecular identifiers (UMIs) are incorporated to facilitate PCR error correction and clonotype quantification during data analysis, and unique dual indexes (UDIs) allow users to multiplex more samples and to provide confidence when sequencing on patterned flow cells. At the end, generated indexed libraries are ready for sequencing on Illumina platforms.

This kit supports up to 48 rxns.

Variable and highly complex samples are accommodated without the need for RNA purification or T-cell enrichment.TRA and TRB libraries were generated from 1, 10, and 100 ng of human CD3+ T‑cell total RNA (Panel A), 1,000 and 10,000 CD3+ T cells (Panel B), and 20 ng of whole-blood RNA extracted from three different samples (Panel C). The sequence reads were processed by Cogent NGS Immune Profiler.

The number of clonotypes detected reveals biological variation.Duplicate libraries were generated from 10 ng RNA extracted from single-donor PBMC samples (P1-P6) and sequenced on an Illumina MiSeq system using 300-bp paired-end reads to obtain approximately 1.5 million reads per sample. Resulting sequencing reads were processed with Cogent NGS Immune Profiler. Clonotype numbers from different donors for TRA (blue) and TRB (orange) are shown. Error bars shown represent the standard error between the duplicates.