The following protocols provide instructions on creating single-cell mRNA libraries with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells. To create the library, cDNA of gene targets are encoded on BD Rhapsody™ Cell Capture Beads. This targeted library approach employs a two-step nested amplification, followed by an index PCR step. The generated library can be sequenced on various Illumina sequencers.
BD Rhapsody VDJ protocol is compatible with BD Rhapsody - Targeted, Abseq and SMK protocols either individually or combined.