The Ion AmpliSeq Mouse BCR IGH SR Assay, RNA, is a robust, targeted next-generation sequencing (NGS) assay for use in basic and translational immunology, immuno-oncology, hemato–oncology, and vaccine research. It is designed to accurately identify and measure the clonal expansion of B lymphocytes in blood, peripheral blood leukocytes (PBLs), peripheral blood mononuclear cells (PBMCs), and fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) samples. The assay identifies unique B-cell clones through targeting of the highly diverse complementarity-determining region 3 (CDR3) of the B-cell receptor (BCR) immunoglobulin heavy (IGH) chain from genomic DNA template . The nucleotide sequence of the IGH CDR3 region serves as a natural barcode to enable clone tracking and measurements of B-cell clonal expansion and diversity. Analysis of IGH CDR3-region amino acid motifs may reveal signatures of B-cell responses to defined antigens.

Benefits of the Ion AmpliSeq Mouse BCR IGH SR Assay, RNA, include:
• Compatibility with a vast array of research sample types, including FFPE tissue, fresh-frozen (FF) tissue, whole blood, PBLs, and PBMCs
• High sensitivity and low limit of detection (LoD) for rare clone identification through dual-barcode indexing
• Efficient workflow with 48 hour sample-to-results time
• Flexible input requirements ranging from 100 ng to 1 μg
• Streamlined and user-friendly informatics solution with with multi-sample analysis functionality

The Ion AmpliSeq Mouse BCR IGH SR Assay, RNA, provides a single pool of multiplex PCR primers and library reagents to generate 90-bp amplicons that can be sequenced on all chip types supported by the Ion GeneStudio S5 sequencing systems, allowing you to pick the best multiplexing configuration for your unique sample batching needs and throughput requirements. The entire workflow, (from library preparation to analysis of samples, can be accomplished in two days using the Ion Chef templating system and Ion GeneStudio S5 system..

The Ion AmpliSeq Mouse BCR IGH SR Assay, RNA, with its high sensitivity and specificity, supports key applications in immunology, immuno-oncology, hemato-oncology, and vaccine research. The high-sensitivity dual-barcode indexing, flexible input requirements (100 ng–1 μg), high-depth sequencing, and high-throughput capability make this assay ideal for testing variety of hypothesis for basic or translational biomarker research. A small fragment-size requirement for the input materials for library creation and high multiplexing on the Ion 530 Chip of the Ion GeneStudio S5 sequencer enables researchers to focus on testing a variety of use cases in mouse models, including the role of T cells in generating immune response to immunotherapies such as checkpoint blockade inhibitors, cancer vaccines, or chimeric antigen receptor (CAR) T-cell therapies.

Note: Information about data analysis using Ion Reporter Software v5.12 can be found in the User Guide below.