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  • SMART-Seq Human BCR (with UMIs)
  • Human BCR profiling kit for Illumina sequencing
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  • Human TCRv2 profiling kit for Illumina sequencing
  • Human TCR profiling kit for Illumina sequencing
  • Human scTCR profiling kit for Illumina sequencing
hBCRv2 tech note Learn more about human BCR profiling
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 scTCR profiling kit for Illumina sequencing
  • Mouse repertoire
    • SMART-Seq Mouse BCR (with UMIs)
    • SMART-Seq Mouse TCR (with UMIs)
    • Mouse BCR profiling kit for Illumina sequencing
    • Mouse TCR profiling kit for Illumina sequencing
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hBCRv2 tech note Learn more about human BCR profiling

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.

 More  Less
Cat. # Product Size Price License Quantity Details
634777 SMART-Seq® Human BCR (with UMIs) 24 Rxns USD $1406.00

License Statement

ID Number  
392 This Product is protected by one or more patents from the family consisting of:US10941397, People's Republic of China Patent: ZL201480057094.4, US10781443, US10954510, DE602014069266.4, EP3058104, FR3058104, UK3058104, JP6602294, SE3058104, CA2923812, and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family. Information on relevant patents and licenses for this product may be found at: https://www.takarabio.com/patents. 
455 This product is sold under license from Becton Dickinson and Co., and may be the subject of U.S. Patent Nos.: 8,835,358; 9,290,809; 9,315,857; 9,708,659; 9,845,502; 10,047,394; 10,059,991; 10,202,646; 10,392,661; 10,619,203; 11,970,737; 12,060,607; and its foreign counterparts.

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

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634777: SMART-Seq Human BCR (with UMIs)

634777: SMART-Seq Human BCR (with UMIs)

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

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs)

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs)

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs). Input RNA or cells are oligo-dT primed using the dT Primer (dark blue). Following oligo dT-priming, SmartScribe Reverse Transcriptase performs first strand cDNA synthesis on input RNA or cells and adds non-templated nucleotides to the 5’ end of each cDNA molecule (XXXXX). Upon reaching the 5’ end of the RNA template, the SMART UMI Oligo anneals to the non-templated nucleotides (XXXXX), incorporating the UMI (yellow) and partial Illumina adapter (light green) complementary to the hBCR PCR1 Universal Forward primer. Following reverse transcription, semi-nested PCR is performed to amplify BCR cDNAs. In PCR1, the hBCR PCR1 Universal Forward primer anneals to the complementary sequence carried by the SMART UMI Oligo (light green), incorporating the Illumina Read2 sequence (dark green). The hBCR PCR1 Reverse primer (orange) anneals to sequences in the constant regions of BCR cDNAs to amplify the entire BCR V(D)J region. During the PCR2, the nested hBCR LC (light chain) or hBCR HC (heavy chain) PCR2 Reverse primers anneal to sequences in BCR constant regions that are internal to the sequences bound by the hBCR PCR1 Reverse primer and add the Illumina Read 1 sequence (light purple). In the same PCR2 reaction, Unique Dual Index Kit primers anneal to the sequence added by hBCR PCR1 Universal Forward primer or the hBCR LC or HC PCR2 Reverse primers to add Illumina P7-i7 and P5-i5 index sequences (dark blue). The result is a sequencing-ready library that contains the entire BCR light chain and/or heavy chain variable regions with a small portion of the constant region.

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 $623.00

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 Image Data

Back

634752: Unique Dual Index Kit (1-96)

634752: Unique Dual Index Kit (1-96)
634753 Unique Dual Index Kit (97–192) 96 Rxns USD $623.00

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 Image Data

Back

634753: Unique Dual Index Kit (97-192)

634753: Unique Dual Index Kit (97-192)
634754 Unique Dual Index Kit (193–288) 96 Rxns USD $623.00

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 Image Data

Back

634754: Unique Dual Index Kit (193-288)

634754: Unique Dual Index Kit (193-288)
634755 Unique Dual Index Kit (289–384) 96 Rxns USD $623.00

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 Image Data

Back

634755: Unique Dual Index Kit (289-384)

634755: Unique Dual Index Kit (289-384)
634756 Unique Dual Index Kit (1–24) 24 Rxns USD $237.00

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 Image Data

Back

634756: Unique Dual Index Kit (1-24)

634756: Unique Dual Index Kit (1-24)
634778 SMART-Seq® Human BCR (with UMIs) 96 Rxns USD $4982.00

License Statement

ID Number  
392 This Product is protected by one or more patents from the family consisting of:US10941397, People's Republic of China Patent: ZL201480057094.4, US10781443, US10954510, DE602014069266.4, EP3058104, FR3058104, UK3058104, JP6602294, SE3058104, CA2923812, and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family. Information on relevant patents and licenses for this product may be found at: https://www.takarabio.com/patents. 
455 This product is sold under license from Becton Dickinson and Co., and may be the subject of U.S. Patent Nos.: 8,835,358; 9,290,809; 9,315,857; 9,708,659; 9,845,502; 10,047,394; 10,059,991; 10,202,646; 10,392,661; 10,619,203; 11,970,737; 12,060,607; and its foreign counterparts.

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

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs)

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs)

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs). Input RNA or cells are oligo-dT primed using the dT Primer (dark blue). Following oligo dT-priming, SmartScribe Reverse Transcriptase performs first strand cDNA synthesis on input RNA or cells and adds non-templated nucleotides to the 5’ end of each cDNA molecule (XXXXX). Upon reaching the 5’ end of the RNA template, the SMART UMI Oligo anneals to the non-templated nucleotides (XXXXX), incorporating the UMI (yellow) and partial Illumina adapter (light green) complementary to the hBCR PCR1 Universal Forward primer. Following reverse transcription, semi-nested PCR is performed to amplify BCR cDNAs. In PCR1, the hBCR PCR1 Universal Forward primer anneals to the complementary sequence carried by the SMART UMI Oligo (light green), incorporating the Illumina Read2 sequence (dark green). The hBCR PCR1 Reverse primer (orange) anneals to sequences in the constant regions of BCR cDNAs to amplify the entire BCR V(D)J region. During the PCR2, the nested hBCR LC (light chain) or hBCR HC (heavy chain) PCR2 Reverse primers anneal to sequences in BCR constant regions that are internal to the sequences bound by the hBCR PCR1 Reverse primer and add the Illumina Read 1 sequence (light purple). In the same PCR2 reaction, Unique Dual Index Kit primers anneal to the sequence added by hBCR PCR1 Universal Forward primer or the hBCR LC or HC PCR2 Reverse primers to add Illumina P7-i7 and P5-i5 index sequences (dark blue). The result is a sequencing-ready library that contains the entire BCR light chain and/or heavy chain variable regions with a small portion of the constant region.

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 $623.00

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 Image Data

Back

634752: Unique Dual Index Kit (1-96)

634752: Unique Dual Index Kit (1-96)
634753 Unique Dual Index Kit (97–192) 96 Rxns USD $623.00

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 Image Data

Back

634753: Unique Dual Index Kit (97-192)

634753: Unique Dual Index Kit (97-192)
634754 Unique Dual Index Kit (193–288) 96 Rxns USD $623.00

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 Image Data

Back

634754: Unique Dual Index Kit (193-288)

634754: Unique Dual Index Kit (193-288)
634755 Unique Dual Index Kit (289–384) 96 Rxns USD $623.00

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 Image Data

Back

634755: Unique Dual Index Kit (289-384)

634755: Unique Dual Index Kit (289-384)
634756 Unique Dual Index Kit (1–24) 24 Rxns USD $237.00

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 Image Data

Back

634756: Unique Dual Index Kit (1-24)

634756: Unique Dual Index Kit (1-24)
634776 SMART-Seq® Human BCR (with UMIs) 4 x 96 Rxns Inquire for Quotation

License Statement

ID Number  
392 This Product is protected by one or more patents from the family consisting of:US10941397, People's Republic of China Patent: ZL201480057094.4, US10781443, US10954510, DE602014069266.4, EP3058104, FR3058104, UK3058104, JP6602294, SE3058104, CA2923812, and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family. Information on relevant patents and licenses for this product may be found at: https://www.takarabio.com/patents. 
455 This product is sold under license from Becton Dickinson and Co., and may be the subject of U.S. Patent Nos.: 8,835,358; 9,290,809; 9,315,857; 9,708,659; 9,845,502; 10,047,394; 10,059,991; 10,202,646; 10,392,661; 10,619,203; 11,970,737; 12,060,607; and its foreign counterparts.
*

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

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs)

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs)

Schematic of technology and workflow for SMART-Seq Human BCR (with UMIs). Input RNA or cells are oligo-dT primed using the dT Primer (dark blue). Following oligo dT-priming, SmartScribe Reverse Transcriptase performs first strand cDNA synthesis on input RNA or cells and adds non-templated nucleotides to the 5’ end of each cDNA molecule (XXXXX). Upon reaching the 5’ end of the RNA template, the SMART UMI Oligo anneals to the non-templated nucleotides (XXXXX), incorporating the UMI (yellow) and partial Illumina adapter (light green) complementary to the hBCR PCR1 Universal Forward primer. Following reverse transcription, semi-nested PCR is performed to amplify BCR cDNAs. In PCR1, the hBCR PCR1 Universal Forward primer anneals to the complementary sequence carried by the SMART UMI Oligo (light green), incorporating the Illumina Read2 sequence (dark green). The hBCR PCR1 Reverse primer (orange) anneals to sequences in the constant regions of BCR cDNAs to amplify the entire BCR V(D)J region. During the PCR2, the nested hBCR LC (light chain) or hBCR HC (heavy chain) PCR2 Reverse primers anneal to sequences in BCR constant regions that are internal to the sequences bound by the hBCR PCR1 Reverse primer and add the Illumina Read 1 sequence (light purple). In the same PCR2 reaction, Unique Dual Index Kit primers anneal to the sequence added by hBCR PCR1 Universal Forward primer or the hBCR LC or HC PCR2 Reverse primers to add Illumina P7-i7 and P5-i5 index sequences (dark blue). The result is a sequencing-ready library that contains the entire BCR light chain and/or heavy chain variable regions with a small portion of the constant region.

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

634776: SMART-Seq Human BCR (with UMIs)

634776: 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 $623.00

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.

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634752: Unique Dual Index Kit (1-96)

634752: Unique Dual Index Kit (1-96)
634753 Unique Dual Index Kit (97–192) 96 Rxns USD $623.00

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.

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634753: Unique Dual Index Kit (97-192)

634753: Unique Dual Index Kit (97-192)
634754 Unique Dual Index Kit (193–288) 96 Rxns USD $623.00

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.

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634754: Unique Dual Index Kit (193-288)

634754: Unique Dual Index Kit (193-288)
634755 Unique Dual Index Kit (289–384) 96 Rxns USD $623.00

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.

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634755: Unique Dual Index Kit (289-384)

634755: Unique Dual Index Kit (289-384)
634756 Unique Dual Index Kit (1–24) 24 Rxns USD $237.00

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.

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634756: Unique Dual Index Kit (1-24)

634756: Unique Dual Index Kit (1-24)

*You must be logged in to a Purchasing Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

Unique Dual Index Kits

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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  • mRNA and cDNA synthesis
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  • cDNA synthesis kits
  • Reverse transcriptases
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  • Purified total RNA and mRNA
  • PCR
  • Most popular polymerases
  • High-yield PCR
  • High-fidelity PCR
  • GC rich PCR
  • PCR master mixes
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  • In-Fusion seamless cloning
  • Competent cells
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