HUMAN IG RNA MULTIPLEX Kit

The HUMAN IG RNA MULTIPLEX kit is designed for the amplification of IG heavy and light chain cDNA libraries using the combination of highly sensitive multiplex PCR and Unique Molecular Identifiers (UMIs). UMIs are introduced along with 1st strand cDNA synthesis and allow for error-free reconstruction of full-length IG heavy and light chain sequence with hypermutations (except for the FR1 region). In addition, UMIs allow for exact quantification of template cDNA molecules, control for input bottlenecks, and accurate normalization of samples of comparison. Gene-specific primers targeting IG heavy chain constant regions facilitate all IGH isotype discrimination including IgM, IgD, IgG3, IgG1, IgA1, IgG2, IgG4, IgE, and IgA2.
The kit includes a set of reagents sufficient to prepare 24 IGH and 24 IGK/L cDNA libraries starting from 24 RNA samples.
Some B cell populations (e.g. plasma cells, plasmablasts) express much higher IG RNA levels compared to memory or naïve B cells. Thus, for some experiments it can be rational to separate these populations before RNA isolation by FACS or magnetic beads.
The kit allows to start with RNA derived from 100 to 100,000 sorted/purified memory/naïve B cells or from 10 to 1,000 plasma cells/plasmablasts, from PBMC (up to 5x105), or from B cell-containing tissues, and produces indexed ready-to-sequence-on-Illumina libraries.

MiXCR is a universal solution for fast and rigorous extraction and analysis of T-cell and B-cell receptor repertoire high-throughput sequencing data.

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