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SMARTer Mouse TCR a/b Profiling Kit—obtain full-length sequences of TCR-alpha and TCR-beta V(D)J variable regions

The SMARTer Mouse TCR a/b Profiling Kit provides a powerful new solution for those seeking to perform T-cell receptor (TCR) repertoire analysis using NGS. The kit employs a 5'-RACE-based approach to capture complete V(D)J variable regions of TCR transcripts, starting from as little as 10 ng to 500 ng of total RNA obtained from mouse spleen, thymus, or PBMCs, or from 1,000 to 10,000 purified T cells. As the name suggests, the kit can be used to generate data for both TCR-alpha and TCR-beta chain diversity, either in the same experiment or separately.

In contrast with TCR profiling methods that involve PCR multiplexing, each round of library amplification utilizes a single primer pair for each TCR subunit, thereby increasing efficiency and reducing the potential for primer-dimer formation. Multiplex PCR-based approaches are known to promote primer-associated, sequence-specific bias; this bias does not exist with our RACE-based approach, as all variable regions are primed from the same constant-region sequence. Using our ~4-hr protocol, libraries produced with the kit are indexed and ready for sequencing on Illumina platforms.

This kit leverages our proven SMART (Switching Mechanism at 5' End of RNA Template) technology along with LNA technology to amplify TCR mRNA sequences in an unbiased manner with remarkable sensitivity.

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Product

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Details

634403

SMARTer® Mouse TCR a/b Profiling Kit

48 Rxns

USD $3225.00

License Statement

ID Number

275

SMART-Seq2 Technology. This product is sold under exclusive license from Ludwig Institute of Cancer Research, Ltd. and is covered by US Patent No. 10266894, Japanese Patent No. 6336080, and European Patent No. 3036336, and pending U.S. patent application and/or pending claims of foreign counterparts. For license information, please contact a Takara Bio USA, Inc. licensing representative by e-mail at licensing@takarabio.com.

The SMARTer Mouse TCR a/b Profiling Kit provides a powerful new solution for those seeking to perform T-cell receptor (TCR) repertoire analysis using NGS.

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.

Components

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Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach. Panel A. Reverse transcription and PCR amplification of TCR subunit mRNA sequences. First-strand cDNA synthesis is primed by the TCR dT Primer and performed by an MMLV-derived reverse transcriptase (RT). Upon reaching the 5′ end of each mRNA molecule, the RT adds non-templated nucleotides to the first-strand cDNA. The SMART-Seq v4 Oligonucleotide contains a sequence that is complementary to the non-templated nucleotides added by the RT, and the oligo hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART-Seq v4 Oligonucleotide as a template for the incorporation of an additional sequence on the end of the first-strand cDNA. Full-length variable regions of TCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (SMART Primer 1) and the constant region(s) of TCR-α and/or TCR-β subunits (TCRa and/or TCRb Mouse Primer 1). A subsequent round of PCR is performed to further amplify variable regions of TCR-α and/or TCR-β subunits and incorporate adapter sequences using a TCR Primer 2 Forward HT Index and a TCRa and/or TCRb Mouse Primer 2 Reverse HT Index. Included in the primers are adapter and index sequences (Read 2 + i7 + P7 and Read 1 + i5 + P5, respectively) that are compatible with the Illumina sequencing platform. Following purification, size selection, and quality analysis, the TCR cDNA library is ready for sequencing. Panel B. Semi-nested PCR approach for amplification of TCR-α and/or TCR-β subunits. The primer pairs used for the first round of amplification capture the entire variable region(s) and most of the constant region(s) of TCR-α and/or TCR-β cDNA. The second round of amplification retains the entire variable region(s) of TCR-α and/or TCR-β cDNA and a smaller portion of the constant region(s). The anticipated size of the final TCR library cDNA (inserts + adapters) is ~700–800 bp.

634404

SMARTer® Mouse TCR a/b Profiling Kit

96 Rxns

USD $5316.00

License Statement

ID Number

275

The SMARTer Mouse TCR a/b Profiling Kit provides a powerful new solution for those seeking to perform T-cell receptor (TCR) repertoire analysis using NGS.

Notice to purchaser

Components

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Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Overview

Complete V(D)J sequence information—obtain full-length sequences for variable regions of TCR mRNA transcripts.

TCR-alpha and TCR-beta—profile diversity for both TCR subunits, either in the same experiment or separately.
No multiplex PCR required—amplify sequences for each TCR subunit with a single primer pair per reaction.
Illumina-ready sequencing libraries—incorporate Illumina adapter and index sequences in a ligation-independent manner, and multiplex up to 96 libraries in a single flow-cell lane.

More Information

Applications

Mouse TCR repertoire analysis (TCR-alpha and TCR-beta 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.

Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2023 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.

SMARTer Mouse TCR a/b Profiling Kit—obtain full-length sequences of TCR-alpha and TCR-beta V(D)J variable regions

Notice to purchaser

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Assessment of TCR diversity in mouse spleen samples with the SMARTer Mouse TCR a/b Profiling Kit

Assessment of TCR-α clonotype diversity in various tissues of transgenic mice of different ages with the SMARTer Mouse TCR a/b Profiling Kit

634403: SMARTer Mouse TCR a/b Profiling Kit

Notice to purchaser

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Assessment of TCR diversity in mouse spleen samples with the SMARTer Mouse TCR a/b Profiling Kit

Assessment of TCR-α clonotype diversity in various tissues of transgenic mice of different ages with the SMARTer Mouse TCR a/b Profiling Kit

634404: SMARTer Mouse TCR a/b Profiling Kit

Overview

More Information

Applications

Additional product information