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SMARTer Human TCR a/b Profiling Kit v2—flexible kit with ultimate sensitivity for understanding the TCR repertoire

NOTE: We recommend the reconfigured versions of the kits below, SMART-Seq Human TCR (with UMIs) (Cat. # 634780, 634781 & 634779).

SMARTer Human TCR a/b Profiling Kit v2 (Cat. # 634478, 634479) will be phased out soon. Contact your local sales representative for more information.

The SMARTer Human TCR a/b Profiling Kit v2 (TCRv2) is powered by robust chemistry that provides unparalleled sensitivity and reproducibility. The kit leverages SMART (Switching Mechanism at 5' end of RNA Template) full-length cDNA synthesis technology and pairs NGS with a 5'-RACE approach to capture the complete V(D)J variable regions of TRA and TRB genes.

NOTE: We recommend the reconfigured versions of the kits below, SMART-Seq Human TCR (with UMIs) (Cat. # 634780, 634781 & 634779).

SMARTer Human TCR a/b Profiling Kit v2 (Cat. # 634478, 634479) will be phased out soon. Contact your local sales representative for more information.

The kit is designed to work with a range of RNA input amounts (RIN ≥8; depending on the sample type) and has been shown to yield high-quality sequencing libraries from as little as 10 ng to 1 µg of total RNA obtained from peripheral blood leukocytes, 20 ng to 200 ng of total RNA obtained from whole blood, 1 ng to 100 ng of total RNA obtained from T cells, or from 1,000 to 10,000 purified, whole T cells. Libraries can be generated to obtain both alpha and beta chain diversity information. This latest profiling kit also includes unique molecular identifiers (UMI), making it possible to remove reads derived from PCR duplicates and sequencing errors, thus ensuring more accurate and reliable results. In addition to the reagents, the TCRv2 kit includes a dual index (UDI) plate to generate Illumina-compatible sequencing libraries.

Once output files are generated with an Illumina sequencer, they can be analyzed using the Cogent NGS Immune Profiler Software. This software enables high-quality TCR profiling analysis, including count and identification of clonotypes and sequence information for V(D)J regions.

Benefits of this kit include:

Use of UMIs to correct for PCR and sequencing errors
Addition of UDIs, allowing for greater confidence in sequencing on a patterned flow cell (such as the NovaSeq™ system) and the ability to pool a greater number of samples
Flexibility to sequence on any Illumina instrument
Full-length V(D)J analysis on the MiSeq™ system
Access to Cogent NGS Immune Profiler Software, an easy-to-use analysis pipeline tool for users of any bioinformatic experience level, for faster data analysis

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Cat. #	Product	Size
634478	SMARTer® Human TCR a/b Profiling Kit v2	12 Rxns	USD $1440.00
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 12 rxns. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Sensitive and reproducible clonotype detection from a broad range of sample types and RNA amounts. 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 CogentIP. Back The Takara Bio TCR profiling kit identifies a wide range of clonotype counts. 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 CogentIP. Clonotype numbers from different donors for TRA (blue) and TRB (orange) are shown. Error bars shown represent the standard error between the duplicates. Back The Takara Bio TCR profiling kit generates data with sensitivity superior to competitors A dramatically higher clonotype number was observed for TRB after downsampling. (TRA results were similar; data not shown.) 5M PBMC cells were split from two different healthy donors for RNA and gDNA extraction. 1.6 g of gDNA (15% of the total amount of extracted gDNA) was used for library preparation according to manufacturer's instructions. 100 ng of RNA (2% of the total amount of extracted RNA) was used for library preparation. Back 634478: SMARTer Human TCR a/b Profiling Kit v2
634479	SMARTer® Human TCR a/b Profiling Kit v2	48 Rxns	USD $3850.00
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. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Sensitive and reproducible clonotype detection from a broad range of sample types and RNA amounts. 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 CogentIP. Back The Takara Bio TCR profiling kit identifies a wide range of clonotype counts. 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 CogentIP. Clonotype numbers from different donors for TRA (blue) and TRB (orange) are shown. Error bars shown represent the standard error between the duplicates. Back The Takara Bio TCR profiling kit generates data with sensitivity superior to competitors A dramatically higher clonotype number was observed for TRB after downsampling. (TRA results were similar; data not shown.) 5M PBMC cells were split from two different healthy donors for RNA and gDNA extraction. 1.6 g of gDNA (15% of the total amount of extracted gDNA) was used for library preparation according to manufacturer's instructions. 100 ng of RNA (2% of the total amount of extracted RNA) was used for library preparation. Back 634479: SMARTer Human TCR a/b Profiling Kit v2

Overview

Compatible with a wide range of sample inputs: 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 from T cells) or purified, whole T cells (1,000 to 10,000 cells)
Simple PCR amplification: a single primer pair for each TCR (alpha or beta) subunit per reaction
UMI-based correction: removal of reads derived from PCR duplicates and sequencing errors
Sensitive and specific clonotype detection: optimized cDNA library generation
UDI implementation: increased multiplexing and confidence for sequencing on high-throughput sequencers
Illumina-ready sequencing libraries: Illumina-compatible index sequences are incorporated for multiplexing up to 192 libraries in a single run
Flexible sequencing options: either full-length V(D)J information on the MiSeq™ system or CDR3 information on all Illumina platforms
Complete workflow: robust chemistry complemented with use of our Cogent NGS Immune Profiler Software* for sequencing data analysis
*The Cogent NGS Immune Profiler Software is free to use with this kit for both academic and for-profit entities

TCRv2 reproducibly detects clonotypes from total RNA and cells

% Jurkat RNA spiked in to 100 ng of PBMC RNA	Total read count (TRA/TRB)	Without UMI collapse			With UMI collapse
% Jurkat RNA spiked in to 100 ng of PBMC RNA	Total read count (TRA/TRB)	# of TRB raw reads	# of reads for TRBV12‑3-TRBJ1‑2	Detected percentage of Jurkat reads	# of detected UMIs	# of UMIs for TRBV12‑3-TRBJ1‑2	Detected percentage of Jurkat UMIs
10.0%	2,500,000	1,565,005	397,179	25.0%	281,280	62,629	22.0%
1.0%	2,500,000	1,422,102	47,160	3.3%	219,776	6,426	2.9%
0.1%	2,500,000	1,366,127	5,412	0.4%	189,580	631	0.33%
0.01%	2,500,000	1,218,025	521	0.043%	196,615	74	0.038%
0.001%	2,500,000	1,331,465	909	0.068%	197,870	6	0.003%
0.0001%	2,500,000	1,409,199	-	0%	124,149	-	0%
0%	2,500,000	1,222,245	-	0%	197,933	-	0%

Table 1. Assessing the sensitivity and reproducibility of the SMARTer approach. Spike-in analysis was performed in replicate on PBMC RNA samples spiked at varying concentrations (10%, 1%, 0.1%, 0.01%, 0.001%, and 0.0001%) with RNA obtained from a homogeneous population of leukemic Jurkat T cells (containing TRBV12-3-TRBJ1-2 clonotypes). TRB CDR3 regions were amplified from 100 ng of total RNA using the TCRv2 kit and sequenced. Reads of 2 x 150 bp were obtained on an Illumina NextSeq® system. The sequencing reads were downsampled to 2.5 M reads. Read results for spike-in concentrations identified as the reliable concentration limit for each criterion (without and with UMI collapse) have data highlighted in gray. Without UMI collapse, PCR duplicates of TRBV12-3 were observed in 0.0010% of the raw reads.

TCRv2 chemistry detects more clonotypes than competitor methods

More Information

Applications

Human TCR repertoire analysis (TCRα and TCRβ subunits)

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Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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SMARTer Human TCR a/b Profiling Kit v2—flexible kit with ultimate sensitivity for understanding the TCR repertoire

Notice to purchaser

Sensitive and reproducible clonotype detection from a broad range of sample types and RNA amounts.

The Takara Bio TCR profiling kit identifies a wide range of clonotype counts.

The Takara Bio TCR profiling kit generates data with sensitivity superior to competitors

634478: SMARTer Human TCR a/b Profiling Kit v2