The SMARTer Human BCR IgG IgM H/K/L Profiling Kit enables users to analyze B-cell receptor (BCR) repertoires from total RNA samples. This kit can generate libraries from 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. This kit may be used for other sample types, but the protocol may need to be modified. The kit can be used to generate data for both heavy and light chains (kappa and lambda) of human immunoglobulin IgG and IgM.
The SMARTer Human BCR IgG IgM 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. It also incorporates unique molecular identifiers (UMI) to facilitate PCR error correction and clonotype quantification during data analysis.
Included in the kit are primers that incorporate Illumina-specific adapter sequences during cDNA amplification. The protocol generates indexed libraries that are ready for sequencing on Illumina platforms.
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
Applications
- Human BCR repertoire analysis (heavy and light [kappa and lambda] chains)
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