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.