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CAT #: 11010081

IGH Gene Clonality Assay MegaKit for ABI Fluorescence Detection

Assay Use

This Research Use Only assay identifies immunoglobulin heavy chain clonality and is useful for the study of identifying clonal B-cell populations and evaluation of new research and methods in malignancy studies.
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Product Details

  • Summary and Explanation of the Test

    BACKGROUND:

    Rearrangements of the antigen receptor genes occur during ontogeny in B and T lymphocytes.  These gene rearrangements generate products that are unique in length and sequence for each cell.  Therefore, polymerase chain reaction (PCR) assays can be used to identify lymphocyte populations derived from a single cell by detecting the unique V-J gene rearrangements present within these antigen receptor loci.1  This PCR assay employs multiple consensus DNA primers that target conserved genetic regions within the immunoglobulin heavy chain gene.  This test is used to detect the vast majority of clonal B-cell malignancies from DNA.  Test products can be analyzed using a variety of detection formats, including gel and capillary electrophoresis.

    Invivoscribe’s Gene Rearrangement and Translocation Assays represent a simple approach to PCR-based clonality testing. These standardized assays were carefully optimized testing positive and negative control samples using multiplex master mixes.

    SUMMARY:

    This test kit includes six (6) master mixes.  Tubes A, B, and C target the framework 1, 2, and 3 regions (respectively) within the variable (VH) region and the joining region of the Ig heavy chain locus.  Tubes D and E target the diversity (DH) and joining (JH) regions.  The last master mix, the Specimen Control Size Ladder, targets multiple genes and generates a series of amplicons of 100, 200, 300, 400, and 600 base pairs (bp) to ensure that the quality and quantity of input DNA is adequate to yield a valid result.

  • Specimen Requirements

    This assay tests extracted and purified genomic DNA (gDNA). Common sources of gDNA include:

    • 5 cc of peripheral blood, bone marrow biopsy or bone marrow aspirate anti-coagulated with heparin or EDTA. Ship at ambient temperature; OR
    • Formalin-fixed paraffin embedded tissue or slides.

References

1. Miller, JE et al. (1999). Molecular Diagnostics. 4:101-117.
2. Van Dongen, JJM et al. (2003). Leukemia. 17:2257-2317.

Disclaimer

This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.

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