b'Gel & Capillary | RUO AssaysGel and Capillary Gel and CapillaryIGH Gene Rearrangement AssaysResearch Use Only (RUO) Assay IGH Gene Rearrangement Assays Research Use Only (RUO) AssayIGH Gene Rearrangement AssaysAssay Use Specimen RequirementsImmunoglobulin heavy chain (IGH) gene clonality assays are usefulThis assay tests extracted and purified genomic DNA (gDNA). Reagentsfor the study of identifying clonal B-cell populations and evaluationof new research and methods in malignancy studies. References Controls Concentration Units in Assay Units in Assay MegaKitSummary and Explanation of the Test1.JE Miller, SS Wilson, DL Jaye, and M Kronenberg. J. Mol. Diag. IVS-0030 Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesIVS-0029 Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesGenomic DNA is amplified using three PCR master mixes that target4: 101-117 (1999).the three conserved framework regions (FR1, FR2, and FR3) of the2. S Tonegawa. Nature 302: 575-581 (1983). IVS-0000 Polyclonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesIGH gene and the joining (J H ) region. These regions flank the unique,Master Mixes Target Units in Assay Units in Assay MegaKithypervariable, antigen-binding, complementarity determiningIGH Framework 1 Framework 1 + J H 1 x 1500 L tube 10 x 1500 L tubesregion 3 (CDR3). All positive and negative DNA controls, as well as an Amplification Control master mix, are included. PCR products can beIGH Framework 2 Framework 2 + J H 1 x 1500 L tube 10 x 1500 L tubesanalyzed by capillary electrophoresis or standard gel electrophoresisIGH Framework 3 Framework 3 + J H 1 x 1500 L tube 10 x 1500 L tubeswith ethidium bromide staining. Clonality is indicated if one or more of the three framework master mixes generates clonal products. Amplification Control HLA-DQa 1 x 1500 L tube 10 x 1500 L tubesBackground Genes encoding immunoglobulin heavy chain (IGH) moleculesOrdering Informationare assembled from multiple polymorphic gene segments that2 Catalog # Products Quantityundergo rearrangement and selection during B-cell development.Rearrangement of these variable (V H ), diversity (D H ), and joining1-101-0010IGH Gene Rearrangement Assay - Gel Detection30 reactions(J H ) genetic segments result in VDJ products of unique length and1-101-0051IGH Gene Rearrangement Assay - ABI Fluorescence Detection30 reactionssequence. 1,2Clonal IGH rearrangements can be rapidly identified through analyses of the size distributions of DNA products amplified1-101-0071 IGH Gene Rearrangement Assay MegaKit - ABI Fluorescence Detection300 reactionsfrom conserved sequences that flank this region. 2For example, DNA 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 proliferationof a single rearranged clone. 1Figure 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.98Invivoscribe 2021|99'