- DNA sequencing
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SMARTer Mouse BCR IgG H/K/L Profiling Kit—capturing complete V(D)J variable regions of BCR transcripts
B cells are essential parts of the adaptive immune response, expressing B-cell receptors (BCRs) on their surface which allow the recognition of unique molecular patterns in pathogens. Understanding the profiles of BCRs (i.e., the diversity of receptors and the clonotypes, defined by expression of specific H, K, and L gene segments) can not only aid in gaining insights into the adaptive immune response in healthy individuals but also in those with a wide range of diseases.
B cells are essential parts of the adaptive immune response, expressing B-cell receptors (BCRs) on their surface which allow the recognition of unique molecular patterns in pathogens. Understanding the profiles of BCRs, (i.e., the diversity of receptors and the clonotypes, defined by expression of specific H, K, and L gene segments) can not only aid in gaining insights into the adaptive immune response in healthy individuals but also those with a wide range of diseases. Accurate determination of the clonotypes and isotypes expressed by the immune system will aid in a complete picture of the B-cell repertoire.
The new SMARTer Mouse BCR IgG H/K/L Profiling Kit pairs 5' RACE with NGS technology to provide a sensitive, accurate, and optimized approach to BCR profiling. While systems that use multiplex PCR allow for the amplification of multiple BCR genes in one reaction, it may prove challenging with regard to sensitivity, specificity, and biases in the amplification of certain sequences, all of which can lead to difficulties in isotype identification. 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. Downstream sequencing of H/K/L chains allows for accurate identification of top clonotypes and reliable assignment of isotype in a majority of cases.
- Input: 10 ng–3 µg of total RNA from spleen, lymph node, PBMCs, and hybridomas
- Sensitive and specific clonotype detection with optimized cDNA library generation
- Accurate amplification of mouse 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
- Mouse BCR repertoire analysis (heavy and light [kappa and lambda] chains)
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.
Specific, accurate amplification for BCR repertoire profiling
The SMARTer Mouse BCR IgG H/K/L Profiling Kit leverages SMART technology and employs a 5' RACE-like approach to capture complete V(D)J variable regions of BCR transcripts. Read the tech note for key technical features and workflow advantages, as well as application data on amplification of heavy and light chains, clonotype detection, and PCR pooling strategies.View data
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