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  • SMART-Seq Human BCR (with UMIs)
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hBCRv2 tech note Learn more about SMART-Seq Human BCR (with UMIs)
Home › Products › Next-generation sequencing › Immune profiling › Human repertoire › SMART-Seq Human BCR (with UMIs)

Immune profiling

  • Human repertoire
    • 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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hBCRv2 tech note Learn more about SMART-Seq Human BCR (with UMIs)

SMART-Seq Human BCR (with UMIs)

SMART-Seq Human BCR (with UMIs) provides a sensitive and reproducible solution for generating high-quality NGS libraries for profiling the human BCR repertoire. 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 all human B-cell receptor (BCR) heavy (IgG/M/D/A/E) and light (IgK/L) chains.

SMART-Seq Human BCR (with UMIs) provides a sensitive and reproducible solution for generating high-quality NGS libraries for profiling the human BCR repertoire. 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 all human B-cell receptor (BCR) heavy (IgG/M/D/A/E) and light (IgK/L) chains.

The kit is designed to work with a range of RNA inputs, from as little as 10 ng to 1 µg of high-integrity (RIN >8) total RNA from PBMCs (10 ng–1 µg), purified B cells (1–100 ng), spleen or bone marrow (10 ng), or whole blood (100 ng).

Streamline your library preparation with provided premixed primer pools and enzyme premix which reduce the number of reagents and steps. The incorporation of unique molecular identifiers (UMIs) within the workflow allows for PCR error correction and clonotype quantification during data analysis. In addition, up to 384 multiplexed Illumina libraries can be created using the Unique Dual Index Kits (Cat. # 634752–56; sold separately). Easily miniaturize to reduce the price-per-reaction or automate for high-throughput workflows.

As 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 BCR 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.

 More  Less
Cat. # Product Size License Quantity Details
634777 SMART-Seq® Human BCR (with UMIs) 24 Rxns USD $1308.00

License Statement

ID Number  
392 This product is protected by U.S. Patent Nos. 10,941,397 and 10,954,510 and European Patent No. 1485394804 and Japanese Patent No. 2016-524004 and corresponding U.S. pending patents and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.

SMART-Seq Human BCR (with UMIs) generates oligo(dT)-primed, full-length mRNA-seq libraries for all BCR heavy (IgG/M/D/E/A) and light (IgK/L) chains. The incorporation of unique molecular identifiers (UMIs) provides greater accuracy for quantitative gene expression analysis across samples while controlling for PCR errors and amplification biases. Up to 384 multiplexed, Illumina-ready sequencing libraries can be obtained using the Unique Dual Index Kits (Cat. # 634752–56). This kit offers an end-to-end solution including cDNA synthesis, library preparation, and data analysis with our free Cogent NGS bioinformatics tools.

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 BCR clonotype detection

Sensitive and reproducible BCR clonotype detection

Sensitive and reproducible clonotype detection from a broad range of RNA amounts. Human BCR heavy chain (IgA/D/E/G/M) and light chain (IgK/L) libraries were generated from 10, 100, and 1,000 ng of total human PBMC RNA using SMART-Seq Human BCR (with UMIs). The sequence reads were processed using Cogent NGS Immune Profiler Software.

Back

Workflow

Workflow
Fig. 1. SMART-Seq Human BCR (with UMIs) library preparation workflow leverages the SMART technology and semi-nested PCR approach. dT primed first-strand cDNA synthesis is 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 UMI Oligo contains complementary sequences to the non-templated nucleotides added by the RT and hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART UMI Oligo as a template for the incorporation of additional sequences at the end of the first-strand cDNA. Full-length variable regions of BCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (hBCR PCR1 Universal Forward) and the constant region(s) of BCR IgA/D/E/G/M/K/L isotypes (hBCR PCR1 Reverse). In a subsequent round of PCR, the variable regions of BCR-IgA/D/E/G/M and/or BCR- IgK/L subunits are further amplified and adapter sequences and indexes incorporated using hBCR PCR2 HC/LC Reverse primers and UDI primers, generating Illumina compatible BCR cDNA libraries for 300 bp (full-length V(D)J) or 150 bp (CDR3 only) paired-end sequencing. Easily analyze your data and obtain tabulated outputs and plots using our free Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer.

Back

Consistency

Consistency
Fig. 2. SMART-Seq Human BCR (with UMIs) shows consistent performance across a variety of inputs. Bar plot showing clonotype counts for human BCR libraries from PBMC RNA (Panel A) and B-cell RNA (Panel B) at various inputs demonstrate a consistent increase in clonotype count with increase in RNA input. Sequencing outputs were down sampled to ~2 million, 6 million and 12 million reads for PBMC RNA and ~1 million, 5 million and 12 million reads for B-cell RNA. Data was processed using Takara Bio’s Immune profiling software (Cogent NGS Immune Profiler). HC=heavy chain, LC=light chain.

Back

Superior data quality

Superior data quality
Fig. 3. Superior data quality with SMART-Seq Human BCR (with UMIs). Significantly higher clonotype counts are observed with the updated SMART-Seq Human BCR (with UMIs) (BCRv2) compared to both the previous version SMARTer Human BCR IgG IgM H/K/L Profiling Kit (BCRv1) and the competitor kit (Company N). Libraries were generated using 10 ng of human PBMC total RNA according to the manufacturer’s instructions and sequenced on the Illumina platform. Panel A. Sequencing outputs were down sampled to ~1 million for all the libraries. BCRv2 generated approximately 550% more clonotypes than previous version. Panel B. Sequencing outputs were down sampled to ~1 million for Company N libraries and 500,000 each for heavy and light chain libraries for BCRv2. In comparison to Company N, BCRv2 generated approximately 9.5K and 22.9K clonotypes for heavy chain (HC) and light chain (LC) isotypes respectively, representing a 127% increase against Company N.

Back

Sensitivity

Sensitivity
Fig. 4. High sensitivity for the identification of low abundance clonotypes. 100pg, 10pg, 1pg and 0.1pg of RNA extracted from the TIB-190 cell line was spiked into 10 ng of PBMC RNA. Panel A. Clone counts at different spike-in levels are listed in the table. Libraries were normalized to 200,000 reads and all counts were measured after UMI-based consensus collapse. Panel B. Calculated correlation between spike-in RNA proportions and detected clonotype frequencies (both axes logarithmically transformed). Linear relationship between spike-in RNA sample (down to 0.001%) and clonotype frequencies observed with SMART-Seq Human BCR (with UMIs).

Back

Performance

Performance
Fig. 5. Highly reproducible BCR profiling with SMART-Seq Human BCR (with UMIs) between replicates and sequencing platforms. BCR profiling libraries from 10 ng of PBMC RNA and 1 ng B-cell RNA were prepared using SMART-Seq Human BCR (with UMIs). Panel A. Technical replicate libraries prepared with 10 ng PBMC RNA sequenced on the Illumina Miseq platform show high clonotype overlap (87%). Panel B. Libraries prepared using 1 ng B-cell RNA sequenced on the Miseq using a 600-cycle V3 cartridge or 300-cycle V2 cartridge as well as on the Miniseq platform show high clonotype overlap. For all libraries, data generated were downsampled to 1,000,000 reads and analyzed using our Cogent NGS Immune Profiler software.

Back

Comparison with previous version of BCR kit

Comparison with previous version of BCR kit
Fig. 6. Comparison with previous version of BCR kit (SMARTer Human BCR IgG IgM H/K/L Profiling Kit)

Back

Reproducibility

Reproducibility
Fig. 7. Correlation analysis for top 100 clonotypes between two biological replicates

Back

634777: SMART-Seq Human BCR (with UMIs)

634777: SMART-Seq Human BCR (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
634778 SMART-Seq® Human BCR (with UMIs) 96 Rxns USD $4656.00

License Statement

ID Number  
392 This product is protected by U.S. Patent Nos. 10,941,397 and 10,954,510 and European Patent No. 1485394804 and Japanese Patent No. 2016-524004 and corresponding U.S. pending patents and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.

SMART-Seq Human BCR (with UMIs) generates oligo(dT)-primed, full-length mRNA-seq libraries for all BCR heavy (IgG/M/D/E/A) and light (IgK/L) chains. The incorporation of unique molecular identifiers (UMIs) provides greater accuracy for quantitative gene expression analysis across samples while controlling for PCR errors and amplification biases. Up to 384 multiplexed, Illumina-ready sequencing libraries can be obtained using the Unique Dual Index Kits (Cat. # 634752–56). This kit offers an end-to-end solution including cDNA synthesis, library preparation, and data analysis with our free Cogent NGS bioinformatics tools.

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 BCR clonotype detection

Sensitive and reproducible BCR clonotype detection

Sensitive and reproducible clonotype detection from a broad range of RNA amounts. Human BCR heavy chain (IgA/D/E/G/M) and light chain (IgK/L) libraries were generated from 10, 100, and 1,000 ng of total human PBMC RNA using SMART-Seq Human BCR (with UMIs). The sequence reads were processed using Cogent NGS Immune Profiler Software.

Back

Workflow

Workflow
Fig. 1. SMART-Seq Human BCR (with UMIs) library preparation workflow leverages the SMART technology and semi-nested PCR approach. dT primed first-strand cDNA synthesis is 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 UMI Oligo contains complementary sequences to the non-templated nucleotides added by the RT and hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART UMI Oligo as a template for the incorporation of additional sequences at the end of the first-strand cDNA. Full-length variable regions of BCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (hBCR PCR1 Universal Forward) and the constant region(s) of BCR IgA/D/E/G/M/K/L isotypes (hBCR PCR1 Reverse). In a subsequent round of PCR, the variable regions of BCR-IgA/D/E/G/M and/or BCR- IgK/L subunits are further amplified and adapter sequences and indexes incorporated using hBCR PCR2 HC/LC Reverse primers and UDI primers, generating Illumina compatible BCR cDNA libraries for 300 bp (full-length V(D)J) or 150 bp (CDR3 only) paired-end sequencing. Easily analyze your data and obtain tabulated outputs and plots using our free Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer.

Back

Consistency

Consistency
Fig. 2. SMART-Seq Human BCR (with UMIs) shows consistent performance across a variety of inputs. Bar plot showing clonotype counts for human BCR libraries from PBMC RNA (Panel A) and B-cell RNA (Panel B) at various inputs demonstrate a consistent increase in clonotype count with increase in RNA input. Sequencing outputs were down sampled to ~2 million, 6 million and 12 million reads for PBMC RNA and ~1 million, 5 million and 12 million reads for B-cell RNA. Data was processed using Takara Bio’s Immune profiling software (Cogent NGS Immune Profiler). HC=heavy chain, LC=light chain.

Back

Superior data quality

Superior data quality
Fig. 3. Superior data quality with SMART-Seq Human BCR (with UMIs). Significantly higher clonotype counts are observed with the updated SMART-Seq Human BCR (with UMIs) (BCRv2) compared to both the previous version SMARTer Human BCR IgG IgM H/K/L Profiling Kit (BCRv1) and the competitor kit (Company N). Libraries were generated using 10 ng of human PBMC total RNA according to the manufacturer’s instructions and sequenced on the Illumina platform. Panel A. Sequencing outputs were down sampled to ~1 million for all the libraries. BCRv2 generated approximately 550% more clonotypes than previous version. Panel B. Sequencing outputs were down sampled to ~1 million for Company N libraries and 500,000 each for heavy and light chain libraries for BCRv2. In comparison to Company N, BCRv2 generated approximately 9.5K and 22.9K clonotypes for heavy chain (HC) and light chain (LC) isotypes respectively, representing a 127% increase against Company N.

Back

Sensitivity

Sensitivity
Fig. 4. High sensitivity for the identification of low abundance clonotypes. 100pg, 10pg, 1pg and 0.1pg of RNA extracted from the TIB-190 cell line was spiked into 10 ng of PBMC RNA. Panel A. Clone counts at different spike-in levels are listed in the table. Libraries were normalized to 200,000 reads and all counts were measured after UMI-based consensus collapse. Panel B. Calculated correlation between spike-in RNA proportions and detected clonotype frequencies (both axes logarithmically transformed). Linear relationship between spike-in RNA sample (down to 0.001%) and clonotype frequencies observed with SMART-Seq Human BCR (with UMIs).

Back

Performance

Performance
Fig. 5. Highly reproducible BCR profiling with SMART-Seq Human BCR (with UMIs) between replicates and sequencing platforms. BCR profiling libraries from 10 ng of PBMC RNA and 1 ng B-cell RNA were prepared using SMART-Seq Human BCR (with UMIs). Panel A. Technical replicate libraries prepared with 10 ng PBMC RNA sequenced on the Illumina Miseq platform show high clonotype overlap (87%). Panel B. Libraries prepared using 1 ng B-cell RNA sequenced on the Miseq using a 600-cycle V3 cartridge or 300-cycle V2 cartridge as well as on the Miniseq platform show high clonotype overlap. For all libraries, data generated were downsampled to 1,000,000 reads and analyzed using our Cogent NGS Immune Profiler software.

Back

Comparison with previous version of BCR kit

Comparison with previous version of BCR kit
Fig. 6. Comparison with previous version of BCR kit (SMARTer Human BCR IgG IgM H/K/L Profiling Kit)

Back

Reproducibility

Reproducibility
Fig. 7. Correlation analysis for top 100 clonotypes between two biological replicates

Back

634778: SMART-Seq Human BCR (with UMIs)

634778: SMART-Seq Human BCR (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
634776 SMART-Seq® Human BCR (with UMIs) 4 x 96 Rxns Inquire for Quotation

License Statement

ID Number  
392 This product is protected by U.S. Patent Nos. 10,941,397 and 10,954,510 and European Patent No. 1485394804 and Japanese Patent No. 2016-524004 and corresponding U.S. pending patents and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.
*

SMART-Seq Human BCR (with UMIs) generates oligo(dT)-primed, full-length mRNA-seq libraries for all BCR heavy (IgG/M/D/E/A) and light (IgK/L) chains. The incorporation of unique molecular identifiers (UMIs) provides greater accuracy for quantitative gene expression analysis across samples while controlling for PCR errors and amplification biases. Up to 384 multiplexed, Illumina-ready sequencing libraries can be obtained using the Unique Dual Index Kits (Cat. # 634752–56). This kit offers an end-to-end solution including cDNA synthesis, library preparation, and data analysis with our free Cogent NGS bioinformatics tools.

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 BCR clonotype detection

Sensitive and reproducible BCR clonotype detection

Sensitive and reproducible clonotype detection from a broad range of RNA amounts. Human BCR heavy chain (IgA/D/E/G/M) and light chain (IgK/L) libraries were generated from 10, 100, and 1,000 ng of total human PBMC RNA using SMART-Seq Human BCR (with UMIs). The sequence reads were processed using Cogent NGS Immune Profiler Software.

Back

Workflow

Workflow
Fig. 1. SMART-Seq Human BCR (with UMIs) library preparation workflow leverages the SMART technology and semi-nested PCR approach. dT primed first-strand cDNA synthesis is 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 UMI Oligo contains complementary sequences to the non-templated nucleotides added by the RT and hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART UMI Oligo as a template for the incorporation of additional sequences at the end of the first-strand cDNA. Full-length variable regions of BCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (hBCR PCR1 Universal Forward) and the constant region(s) of BCR IgA/D/E/G/M/K/L isotypes (hBCR PCR1 Reverse). In a subsequent round of PCR, the variable regions of BCR-IgA/D/E/G/M and/or BCR- IgK/L subunits are further amplified and adapter sequences and indexes incorporated using hBCR PCR2 HC/LC Reverse primers and UDI primers, generating Illumina compatible BCR cDNA libraries for 300 bp (full-length V(D)J) or 150 bp (CDR3 only) paired-end sequencing. Easily analyze your data and obtain tabulated outputs and plots using our free Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer.

Back

Consistency

Consistency
Fig. 2. SMART-Seq Human BCR (with UMIs) shows consistent performance across a variety of inputs. Bar plot showing clonotype counts for human BCR libraries from PBMC RNA (Panel A) and B-cell RNA (Panel B) at various inputs demonstrate a consistent increase in clonotype count with increase in RNA input. Sequencing outputs were down sampled to ~2 million, 6 million and 12 million reads for PBMC RNA and ~1 million, 5 million and 12 million reads for B-cell RNA. Data was processed using Takara Bio’s Immune profiling software (Cogent NGS Immune Profiler). HC=heavy chain, LC=light chain.

Back

Superior data quality

Superior data quality
Fig. 3. Superior data quality with SMART-Seq Human BCR (with UMIs). Significantly higher clonotype counts are observed with the updated SMART-Seq Human BCR (with UMIs) (BCRv2) compared to both the previous version SMARTer Human BCR IgG IgM H/K/L Profiling Kit (BCRv1) and the competitor kit (Company N). Libraries were generated using 10 ng of human PBMC total RNA according to the manufacturer’s instructions and sequenced on the Illumina platform. Panel A. Sequencing outputs were down sampled to ~1 million for all the libraries. BCRv2 generated approximately 550% more clonotypes than previous version. Panel B. Sequencing outputs were down sampled to ~1 million for Company N libraries and 500,000 each for heavy and light chain libraries for BCRv2. In comparison to Company N, BCRv2 generated approximately 9.5K and 22.9K clonotypes for heavy chain (HC) and light chain (LC) isotypes respectively, representing a 127% increase against Company N.

Back

Sensitivity

Sensitivity
Fig. 4. High sensitivity for the identification of low abundance clonotypes. 100pg, 10pg, 1pg and 0.1pg of RNA extracted from the TIB-190 cell line was spiked into 10 ng of PBMC RNA. Panel A. Clone counts at different spike-in levels are listed in the table. Libraries were normalized to 200,000 reads and all counts were measured after UMI-based consensus collapse. Panel B. Calculated correlation between spike-in RNA proportions and detected clonotype frequencies (both axes logarithmically transformed). Linear relationship between spike-in RNA sample (down to 0.001%) and clonotype frequencies observed with SMART-Seq Human BCR (with UMIs).

Back

Performance

Performance
Fig. 5. Highly reproducible BCR profiling with SMART-Seq Human BCR (with UMIs) between replicates and sequencing platforms. BCR profiling libraries from 10 ng of PBMC RNA and 1 ng B-cell RNA were prepared using SMART-Seq Human BCR (with UMIs). Panel A. Technical replicate libraries prepared with 10 ng PBMC RNA sequenced on the Illumina Miseq platform show high clonotype overlap (87%). Panel B. Libraries prepared using 1 ng B-cell RNA sequenced on the Miseq using a 600-cycle V3 cartridge or 300-cycle V2 cartridge as well as on the Miniseq platform show high clonotype overlap. For all libraries, data generated were downsampled to 1,000,000 reads and analyzed using our Cogent NGS Immune Profiler software.

Back

Comparison with previous version of BCR kit

Comparison with previous version of BCR kit
Fig. 6. Comparison with previous version of BCR kit (SMARTer Human BCR IgG IgM H/K/L Profiling Kit)

Back

Reproducibility

Reproducibility
Fig. 7. Correlation analysis for top 100 clonotypes between two biological replicates

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

Unique Dual Index Kits
Cogent NGS Immune Profiler
Cogent NGS Immune Viewer

Overview

  • Complete BCR repertoire profiling: includes all heavy (IgG, IgM, IgA, IgD, IgE) and light (IgK, IgL) BCR chains
  • Supports various inputs: high-integrity RNA from PBMCs (10 ng–1 µg), B cells (1–100 ng), spleen or bone marrow (10 ng), and whole blood (100 ng)
  • Streamlined workflow: primer pools for heavy and light immunoglobulin chains and enzyme premixes provided
  • Increased accuracy: UMI-based analysis controls for PCR duplicates and errors, correcting for artifacts in clonotype diversity and abundance assessments
  • Flexible: sequence either the full-length V(D)J region or the CDR3 only and multiplex with up to 384 UDIs
  • Economical and high-throughput friendly: can be miniaturized to reduce costs, or automated
  • End-to-end solution: pair with free-to-use Cogent NGS Immune Profiler Software and Cogent NGS Immune Viewer for data analysis and visualization

Achieve sensitive and reproducible BCR clonotype detection

Figure 1. Sensitive and reproducible clonotype detection from a broad range of RNA amounts. Human BCR heavy chain (IgA/D/E/G/M) and light chain (IgK/L) libraries were generated from 10, 100, and 1,000 ng of total human PBMC RNA using SMART-Seq Human BCR (with UMIs). The sequence reads were processed using Cogent NGS Immune Profiler Software.


More Information

Applications 

Complete human BCR repertoire (IgG/A/M/D/E/K/L) analysis. 

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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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.

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES (EXCEPT AS SPECIFICALLY NOTED).

Clontech, TaKaRa, cellartis

  • Products
  • COVID-19 research
  • Next-generation sequencing
  • Diagnostic solutions
  • Real-time PCR
  • Stem cell research
  • mRNA and cDNA synthesis
  • PCR
  • Cloning
  • Nucleic acid purification
  • Gene function
  • Protein research
  • Antibodies and ELISA
  • New products
  • Special offers
  • COVID-19 research
  • Viral detection with qPCR
  • SARS-CoV-2 pseudovirus
  • Human ACE2 stable cell line
  • Viral RNA isolation
  • Viral and host sequencing
  • Vaccine development
  • CRISPR screening
  • Drug discovery
  • Immune profiling
  • Publications
  • Next-generation sequencing
  • RNA-seq
  • DNA-seq
  • Single-cell NGS automation
  • Reproductive health
  • Bioinformatics tools
  • Whole genome amplification
  • Immune profiling
  • Diagnostic solutions
  • Reproductive health
  • Real-time PCR
  • Real-time PCR kits
  • Reverse transcription prior to qPCR
  • High-throughput qPCR solutions
  • RNA extraction and analysis for real-time qPCR
  • Stem cell research
  • Media and supplements
  • Stem cells and stem cell-derived cells
  • Single-cell cloning of edited hiPS cells
  • mRNA and cDNA synthesis
  • In vitro transcription
  • cDNA synthesis kits
  • Reverse transcriptases
  • RACE kits
  • Purified cDNA & genomic DNA
  • Purified total RNA and mRNA
  • PCR
  • Most popular polymerases
  • High-yield PCR
  • High-fidelity PCR
  • GC rich PCR
  • PCR master mixes
  • Cloning
  • In-Fusion seamless cloning
  • Competent cells
  • Ligation kits
  • Restriction enzymes
  • Nucleic acid purification
  • Plasmid purification kits
  • Genomic DNA purification kits
  • DNA cleanup kits
  • RNA purification kits
  • Cell-free DNA purification kits
  • Microbiome
  • Gene function
  • Gene editing
  • Viral transduction
  • Fluorescent proteins
  • T-cell transduction and culture
  • Tet-inducible expression systems
  • Transfection reagents
  • Cell biology assays
  • Protein research
  • Purification products
  • Two-hybrid and one-hybrid systems
  • Mass spectrometry reagents
  • Antibodies and ELISA
  • Primary antibodies and ELISAs by research area
  • Fluorescent protein antibodies
  • Special offers
  • Free samples
  • TB Green qPCR sale
  • PrimeSTAR enzyme promo
  • Try BcaBEST DNA Polymerase ver.2.0
  • RNA purification sale
  • Capturem IP and Co-IP sale
  • Baculovirus titration kits early access program
  • NGS bundle and save
  • Free sample: PrimePath Direct Saliva SARS-CoV-2 Detection Kit
  • TALON his-tag purification resin special offer
  • GoStix Plus special offers
  • PCR samples
  • Services & Support
  • Instrument services
  • Stem cell services
  • Gene and cell therapy manufacturing
  • Customer service
  • Sales
  • Shipping & delivery
  • Technical support
  • Feedback
  • Online tools
  • Partnering & Licensing
  • Vector information
  • Instrument services
  • Apollo services
  • ICELL8 services
  • SmartChip services
  • Stem cell services
  • Clinical-grade stem cell services
  • Research-grade stem cell services
  • Outsourcing stem cell-based disease model development
  • Gene and cell therapy manufacturing
  • Services
  • Facilities
  • Our process
  • Resources
  • Sales
  • Make an appointment with your sales rep
  • Online tools
  • GoStix Plus FAQs
  • Vector information
  • Vector document overview
  • Vector document finder
  • Learning centers
  • Automation systems
  • Next-generation sequencing
  • cDNA synthesis
  • Real-time PCR
  • Nucleic acid purification
  • PCR
  • Cloning
  • Stem cell research
  • Gene function
  • Protein research
  • Antibodies and ELISA
  • Automation systems
  • SmartChip Real-Time PCR System introduction
  • ICELL8 introduction
  • Next-generation sequencing
  • Technical notes
  • Featured kits
  • Technology and application overviews
  • FAQs and tips
  • DNA-seq protocols
  • Bioinformatics resources
  • Webinars
  • Real-time PCR
  • Overview
  • Reaction size guidelines
  • Guest webinar: extraction-free SARS-CoV-2 detection
  • Guest webinar: developing and validating molecular diagnostic tests
  • Technical notes
  • Nucleic acid purification
  • Nucleic acid extraction webinars
  • Product demonstration videos
  • Product finder
  • Plasmid kit selection guide
  • RNA purification kit finder
  • PCR
  • Citations
  • Selection guides
  • Technical notes
  • FAQ
  • Cloning
  • In-Fusion Cloning general information
  • Primer design and other tools
  • In‑Fusion Cloning tips and FAQs
  • Applications and technical notes
  • Stem cell research
  • Overview
  • Protocols
  • Technical notes
  • Gene function
  • Gene editing
  • Viral transduction
  • T-cell transduction and culture
  • Inducible systems
  • Cell biology assays
  • Protein research
  • Capturem technology
  • Antibody immunoprecipitation
  • His-tag purification
  • Other tag purification
  • Expression systems
  • APPLICATIONS
  • Molecular diagnostics
  • Vaccine development
  • Pathogen detection
  • Immunotherapy research
  • Cancer research
  • Alzheimer's disease research
  • Reproductive health technologies
  • Molecular diagnostics
  • Interview: adapting to change with Takara Bio
  • Applications
  • Solutions
  • Partnering
  • Webinar: Speeding up diagnostic development
  • Contact us
  • Vaccine development
  • Characterizing the viral genome and host response
  • Identifying and cloning vaccine targets
  • Expressing and purifying vaccine targets
  • Immunizing mice and optimizing vaccine targets
  • Pathogen detection
  • Sample prep
  • Detection methods
  • Identification and characterization
  • SARS-CoV-2
  • Antibiotic-resistant bacteria
  • Food crop pathogens
  • Waterborne disease outbreaks
  • Viral-induced cancer
  • Immunotherapy research
  • T-cell therapy
  • Antibody therapeutics
  • T-cell receptor profiling
  • TBI initiatives in cancer therapy
  • Cancer research
  • Sample prep from FFPE tissue
  • Sample prep from plasma
  • Cancer biomarker discovery
  • Cancer biomarker quantification
  • Single cancer cell analysis
  • Cancer genomics and epigenomics
  • HLA typing in cancer
  • Gene editing for cancer therapy/drug discovery
  • Alzheimer's disease research
  • Antibody engineering
  • Sample prep from FFPE tissue
  • Single-cell sequencing
  • Reproductive health technologies
  • Preimplantation genetic testing
  • ESM Collection Kit forms
  • About
  • BioView blog
  • That's Good Science!
  • Our brands
  • Our history
  • Announcements
  • Events
  • Careers
  • Trademarks
  • License statements
  • Quality and compliance
  • Takara Bio affiliates & distributors
  • Need help?
  • Website FAQs
  • BioView blog
  • Automation
  • Cancer research
  • Career spotlights
  • Current events
  • Customer stories
  • Gene editing
  • Research news
  • Single-cell analysis
  • Stem cell research
  • Tips and troubleshooting
  • Women in STEM
  • That's Good Support!
  • About our blog
  • That's Good Science!
  • SMART-Seq Pro Biomarker Discovery Contest
  • DNA extraction educational activity
  • That's Good Science Podcast
  • Season one
  • Season two
  • Season three
  • Our brands
  • Takara
  • Clontech
  • Cellartis
  • Events
  • Biomarker discovery events
  • Calendar
  • Conferences
  • Speak with us
  • Careers
  • Company benefits
  • Takara Bio affiliates & distributors
  • United States and Canada
  • China
  • Japan
  • Korea
  • Europe
  • India
  • Affiliates & distributors, by country
  • Need help?
  • Privacy request
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