The Oncomine BCR IGH-SR Assay, RNA, is a robust, targeted next-generation sequencing (NGS) assay designed for hematology-oncology and minimal residual disease (MRD) monitoring research, pharmacodynamics and biomarker analysis of immunotherapy, as well as infectious and autoimmune disease research. The assay kit includes a single pool of multiplex PCR primers, total RNA-to-cDNA synthesis kit and Ion AmpliSeq library reagents.
The assay accurately identifies and measures clonal expansion of B lymphocytes using total RNA from bone marrow, whole blood, peripheral blood leukocytes (PBLs), peripheral blood mononuclear cells (PBMCs), and fresh-frozen and formalin-fixed paraffin-embedded (FFPE) research samples. The assay identifies unique B-cell clones through targeting of the highly diverse complementarity-determining region 3 (CDR3) of the B-cell receptor IGH chain from mRNA template via AmpliSeq multiplex framework 3 and joining gene primers. The nucleotide sequence of the IGH CDR3 region serves as a natural barcode to enable clone tracking for MRD clinical research and measurements of B-cell clonal expansion and diversity for pharmacodynamics and vaccine biomarker research. IGH CDR3 region amino acid motifs may reveal signatures of B-cell responses to defined antigens for use as future markers of autoimmune or infectious disease. RNA input improves detection of changes in plasmablast and plasma cell populations following immune challenge or administration of immunomodulatory agents. For DNA samples, please see Cat. No. A45483.
Benefits of the Oncomine BCR IGH SR Assay, RNA, include: • High sensitivity and exceptionally low limit of detection for rare clone detection and tracking in research samples via dual-barcode indexing and high template capture efficiency of AmpliSeq multiplex PCR • Compatibility with an array of research sample types, including fresh-frozen and FFPE tissue, bone marrow, whole blood, PBLs, and PBMCs • Assay primers design to amplify all variable and joining gene alleles in the gold-standard IMGT database • Ability to generate large libraries with minimal input requirement, facilitating detection of rare clones at a frequency of 10-6 from a single library preparation • Efficient workflow using the Ion Chef templating system and Ion GeneStudio S5 sequencing system, enabling library-to-results time within 48 hours • Flexible input requirements ranging from 25 ng to 2 µg • Streamlined and user-friendly informatics solution with automated clonotyping, clonal lineage analysis, reporting of key repertoire features, and multi-sample analysis capability to enable longitudinal tracking in research samples • Ultra-low substitution error rate minimizing artifacts arising from sequencing errors, enabling highly accurate assessment of clonal heterogeneity in malignant B-cell research samples of interest
The Oncomine BCR IGH-SR Assay, RNA, uses multiplex PCR primers to generate 110-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.