b'Gel & Capillary | RUO AssaysGel and CapillaryIGH Gene Rearrangement AssaysResearch Use Only (RUO) AssayIGH Gene Rearrangement AssaysAssay Use Background IGH Gene Rearrangement Assays are useful for studies involving: Genes encoding immunoglobulin heavy chain (IGH) molecules are assembled from multiple polymorphic gene segments that Identification of clonal B-cell populations highly suggestive undergo rearrangement and selection during B-cell development. 2 of B-cell malignancies Rearrangement of these variable (V H ), diversity (D H ), and joining Lineage determination of leukemias and lymphomas (J H ) genetic segments result in VDJ products of unique length and sequence. 1,2Clonal IGH rearrangements can be rapidly identified Monitoring and evaluation of disease recurrence through analyses of the size distributions of DNA products amplified Detection and assessment of residual disease from conserved sequences that flank this region. 2For example, DNA Evaluation of new research and methods in malignancy studies isolated from a normal polyclonal population of B cells producesa Gaussian distribution (bell-shaped size curve) of amplified products; whereas, DNA amplified from a clonal B-cell population generates one or two product(s) of unique size that reflect proliferationSummary and Explanation of the Test1of a single rearranged clone. In comparison, southern blot analysis Genomic DNA is amplified using three PCR master mixes that targetrequires 1-2 weeks, is significantly less sensitive, and requires the three conserved framework regions (FR1, FR2, and FR3) of theapproximately one hundred times more DNA than PCR-based assays, IGH gene and the joining (J H ) region. These regions flank the unique,which can be completed in 4-5 hours. 1In addition, tests of samples hypervariable, antigen-binding, complementarity determiningpreviously designated Quantity Not Sufficient (QNS), such as formalin-region 3 (CDR3). All positive and negative DNA controls, as well as anfixed, paraffin embedded (FFPE) tissue sections, routinely produceAmplification Control master mix, are included. The limit of detectiona valid result with PCR methods.of this assay is one clonal B cell in a background of a hundred normal cells. PCR products can be analyzed by capillary electrophoresisor standard gel electrophoresis with ethidium bromide staining. Specimen RequirementsClonality is indicated if one or more of the three framework master 5 mL of peripheral blood, bone marrow biopsy, or bone marrow mixes generates clonal products.aspirate anti-coagulated with heparin or EDTA; or,Minimum 5 mm cube of tissue; or,2 g of genomic DNA; or,Formalin-fixed, paraffin-embedded tissue or slides.Reference1.JE Miller, SS Wilson, DL Jaye, and M Kronenberg. J. Mol. Diag. 4: 101-117 (1999).2. S Tonegawa. Nature 302: 575-581 (1983).Figure Legend: Genomic organization of a rearranged immunoglobulin heavy chain gene on chromosome 14. The blue and green arrows represent primers targeting the conserved framework regions within the variable region gene. The relative location, size range of valid products, and colors correspond to the products generated from each of these regions when differential fluorescence detection methods are used.98'