SMARTer Human BCR IgG IgM H/K/L Profiling Kit—capturing complete V(D)J variable regions of BCR transcripts
The new SMARTer Human BCR IgG IgM H/K/L Profiling Kit pairs 5' RACE with NGS technology to provide a sensitive, accurate, and optimized approach to BCR profiling from RNA input samples. The 5' RACE method reduces variability and allows for priming from the constant region of BCR heavy or light chains. This kit combines these benefits with gene-specific amplification to capture complete V(D)J variable regions of BCR transcripts and provide a highly sensitive and reproducible method for profiling B-cell repertoires.
The new SMARTer Human BCR IgG IgM H/K/L Profiling Kit pairs 5' RACE with NGS technology to provide a sensitive, accurate, and optimized approach to BCR profiling from RNA input samples. The 5' RACE method reduces variability and allows for priming from the constant region of BCR heavy or light chains. This kit combines these benefits with gene-specific amplification to capture complete V(D)J variable regions of BCR transcripts and provide a highly sensitive and reproducible method for profiling B-cell repertoires.
The kit is designed to work with a range of RNA input amounts depending on the sample type, and has been shown to generate high-quality, Illumina-ready libraries from as little as 10 ng to 1 µg of total RNA from peripheral blood mononuclear cells (PBMCs) and 1 to 100 ng of total RNA from B cells. Data can be generated for both heavy and light chains (kappa and lambda) of human immunoglobulin IgG and IgM. This latest profiling kit also includes unique molecular identifiers (UMI), making it possible to remove reads derived from PCR errors or duplication and/or sequencing errors, thus ensuring more accurate and reliable results
Upon completion of a sequencing run, data can be analyzed with our Takara Bio Immune Profiler Software, which uses UMI-based error correction and features MIGEC (Shugay, 2014) and MiXCR software (Bolotin, 2015). This software enables high-quality BCR profiling analysis, including clonotype count and identification and sequence information for V(D)J regions.
Overview
- Input: 10 ng to 1 µg of total RNA from peripheral blood mononuclear cells (PBMCs) or 1 to 100 ng of total RNA from B cells
- UMI-based correction for removal of reads derived from PCR error or duplication and/or sequencing errors
- Sensitive and specific clonotype detection with optimized cDNA library generation
- Accurate amplification of human IgG subclasses and identification via sequencing in a majority of cases
- Optimized PCR pooling strategy for highly sensitive detection of different chains from the same sample
- Flexible pooling strategies accommodate different experimental requirements regarding alignment and identification of primary chain sequences
- Complete workflow with the use of our Takara Bio Immune Profiler Software for sequencing data analysis
- The Takara Bio Immune Profiler Software is free to use, but there may be additional costs associated with underlying third-party software for commercial entities
More Information
Applications
- Human BCR repertoire analysis (heavy and light [kappa and lambda] chains)
References
Shugay M. et al. Towards error-free profiling of immune repertoires. Nat. Methods 11, 653–655 (2014).
Bolotin, D. A. et al. MiXCR: software for comprehensive adaptive immunity profiling. Nat. Methods 12, 380–381 (2015).
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.
Profiling human B-cell receptors with a 5' RACE approach and SMART technology
The SMARTer Human BCR IgG IgM H/K/L Profiling Kit captures entire V(D)J variable regions of BCR transcripts, allowing you to reach a complete and accurate view of human BCR repertoires. Incorporation of UMIs further improves accuracy and reliability by enabling the removal of reads derived from PCR errors or duplication and/or sequencing errors.
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