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  • SMART-Seq Human BCR (with UMIs)
  • Human BCR profiling kit for Illumina sequencing
  • SMART-Seq Human TCR (with UMIs)
  • Human TCRv2 profiling kit for Illumina sequencing
  • Human TCR profiling kit for Illumina sequencing
  • Human scTCR profiling kit for Illumina sequencing
Immune profiling t cell repertoire TCR-seq methods: strengths, weaknesses, and rankings
SMARTer TCRv2 technical note View data generated with the TCRv2 kit
TCR and BCR profiling tips 4 factors to consider for TCR/BCR profiling
Using UMTs in NGS experiments Blog post: Using UMIs in NGS experiments
Home › Products › Next-generation sequencing › Immune profiling › Human repertoire › SMART-Seq Human TCR (with UMIs)

Immune profiling

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    • SMART-Seq Human BCR (with UMIs)
    • Human BCR profiling kit for Illumina sequencing
    • SMART-Seq Human TCR (with UMIs)
    • Human TCRv2 profiling kit for Illumina sequencing
    • Human TCR profiling kit for Illumina sequencing
    • Human scTCR profiling kit for Illumina sequencing
  • Mouse repertoire
    • Mouse BCR profiling kit for Illumina sequencing
    • Mouse TCR profiling kit for Illumina sequencing
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Immune profiling t cell repertoire TCR-seq methods: strengths, weaknesses, and rankings
SMARTer TCRv2 technical note View data generated with the TCRv2 kit
TCR and BCR profiling tips 4 factors to consider for TCR/BCR profiling
Using UMTs in NGS experiments Blog post: Using UMIs in NGS experiments

SMART-Seq Human TCR (with UMIs)

NGS TCR Profiling

NOTE: SMART-Seq Human TCR (with UMIs) is an equivalent replacement for the SMARTer Human TCR a/b Profiling Kit v2 with minor updates. (See a complete list of kits with new names and the existing kits they will replace here). The update (listed below) does not in any way impact the protocols or functional performance of these kits.

For added flexibility, indexes are not included in the kit, but a choice of indexing primers is sold separately (see Unique Dual Index [UDI] kits)

SMART-Seq Human TCR (with UMIs) 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 human TRA and TRB genes.

NOTE: SMART-Seq Human TCR (with UMIs) is an equivalent replacement for the SMARTer Human TCR a/b Profiling Kit v2 with minor updates. (See a complete list of kits with new names and the existing kits they will replace here). The update (listed below) does not in any way impact the protocols or functional performance of these kits.

For added flexibility, indexes are not included in the kit, but a choice of indexing primers is sold separately (see Unique Dual Index [UDI] kits)

SMART-Seq Human TCR (with UMIs) 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 human TRA and TRB genes.

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. Generate up to 384 multiplexed Illumina libraries using the Unique Dual Index kits (Cat. Nos. 634752–634756; sold separately).

As a part of a complete solution, analyze your sequencing data with our free-to-use Cogent NGS Immune Profiler Software (Cogent IP). This software enables high-quality TCR profiling analysis, including count and identification of clonotypes and sequence information for V(D)J regions. As a bonus, our cloud-based Cogent NGS Immune Viewer allows you to quickly and easily visualize and produce publication-quality tabulated outputs and figures of your data using output files from Cogent IP.

Benefits of this kit include:

  • Use of UMIs to correct for PCR and sequencing errors
  • Compatibility with 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 or CDR3-only analysis
  • Access to Cogent NGS Immune Profiler Software and Immune Viewer Software, an easy-to-use analysis pipeline for users of any bioinformatic experience level, for faster data analysis
 More  Less
Cat. # Product Size License Quantity Details
634779 SMART-Seq® Human TCR (with UMIs) 4 x 96 Rxns Inquire for Quotation *

SMART-Seq Human TCR (with UMIs) 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 provide confidence when sequencing on patterned flow cells. The kit generates indexed libraries ready for sequencing on Illumina platforms. Easily analyze and visualize your data with Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer tool.

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 You May Also Like Image Data

Back

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

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 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

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.

Required Products

Cat. # Product Size Price License Quantity Details
634752 Unique Dual Index Kit (1–96) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (1–96) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634753 Unique Dual Index Kit (97–192) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (97–192) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634754 Unique Dual Index Kit (193–288) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (193–288) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634755 Unique Dual Index Kit (289–384) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (289–384) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634756 Unique Dual Index Kit (1–24) 24 Rxns USD $215.00

The Unique Dual Index (UDI) Kit (1–24) contains 24 pairs of unique dual-indexed PCR primers, which are a subset of the Unique Dual Index Kit (1–96), Cat. No. 634752. They can be used to generate up to 24 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634780 SMART-Seq® Human TCR (with UMIs) 24 Rxns USD $1308.00

SMART-Seq Human TCR (with UMIs) 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 provide confidence when sequencing on patterned flow cells. The kit generates indexed libraries ready for sequencing on Illumina platforms. Easily analyze and visualize your data with Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer tool.

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 You May Also Like Image Data

Back

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

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 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

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

634780: SMART-Seq Human TCR (with UMIs)

634780: SMART-Seq Human TCR (with UMIs)

Required Products

Cat. # Product Size Price License Quantity Details
634752 Unique Dual Index Kit (1–96) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (1–96) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634753 Unique Dual Index Kit (97–192) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (97–192) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634754 Unique Dual Index Kit (193–288) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (193–288) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634755 Unique Dual Index Kit (289–384) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (289–384) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634756 Unique Dual Index Kit (1–24) 24 Rxns USD $215.00

The Unique Dual Index (UDI) Kit (1–24) contains 24 pairs of unique dual-indexed PCR primers, which are a subset of the Unique Dual Index Kit (1–96), Cat. No. 634752. They can be used to generate up to 24 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634781 SMART-Seq® Human TCR (with UMIs) 96 Rxns USD $4656.00

SMART-Seq Human TCR (with UMIs) 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 provide confidence when sequencing on patterned flow cells. The kit generates indexed libraries ready for sequencing on Illumina platforms. Easily analyze and visualize your data with Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer tool.

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 You May Also Like Image Data

Back

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

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 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

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

634781: SMART-Seq Human TCR (with UMIs)

634781: SMART-Seq Human TCR (with UMIs)

Required Products

Cat. # Product Size Price License Quantity Details
634752 Unique Dual Index Kit (1–96) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (1–96) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634753 Unique Dual Index Kit (97–192) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (97–192) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634754 Unique Dual Index Kit (193–288) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (193–288) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634755 Unique Dual Index Kit (289–384) 96 Rxns USD $579.00

The Unique Dual Index (UDI) Kit (289–384) contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components
634756 Unique Dual Index Kit (1–24) 24 Rxns USD $215.00

The Unique Dual Index (UDI) Kit (1–24) contains 24 pairs of unique dual-indexed PCR primers, which are a subset of the Unique Dual Index Kit (1–96), Cat. No. 634752. They can be used to generate up to 24 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

Documents Components

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 384 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 and web-based Cogent Immune Viewer* for sequencing data analysis and visualization
    *The Cogent NGS Software tools are free to use with this kit for both academic and for-profit entities

TCRv2 reproducibly detects clonotypes from total RNA and cells

Figure 1. Sensitive and reproducible clonotype detection from a broad range of RNA amounts. TRA and TRB libraries were generated from 1, 10, and 100 ng of human CD3+ T-cell total RNA and 1,000 and 10,000 CD3+ T cells. The sequence reads were processed by the Cogent NGS Immune Profiler Software.

% Jurkat RNA spiked in to 100 ng of PBMC RNA Total read count (TRA/TRB) Without UMI collapse With UMI collapse
# 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 SMART-Seq 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 SMARTer Human TCR a/b Profiling Kit v2 (TCRv2) 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

Figure 2. Superior sensitivity and reproducibility. We split 5 M PBMC cells from two different healthy donors for RNA and gDNA extraction. 1.6 µg of gDNA was used for library preparation according to the manufacturer's instructions (15% of the total amount of extracted gDNA). 100 ng of RNA was used for library preparation (2% of the total amount of extracted RNA). Clonotype numbers for TCRa/b libraries are shown from each company (NT: not tested). In the comparison, the Takara Bio TCR profiling kit generated an average of 48.7K and 163K clonotypes for TRA and TRB, respectively, representing a 290% increase against Company Q's RNA-based approach and a 145% increase against Company A's gDNA-based approach. Importantly, the RNA methods used only 2% of the total RNA from the 5 M PBMCs.

More Information

Applications

Human TCR repertoire analysis (TCRα and TCRβ subunits)

Additional product information

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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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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