b'Gel & Capillary | RUO AssaysGel and CapillaryT-Cell Receptor Gamma Gene Research Use Only (RUO) AssayRearrangement AssaysT-Cell Receptor Gamma GeneRearrangement AssaysAssay Use Background T-Cell Receptor Gamma Gene Rearrangement Assays are useful forThe T-cell receptor gamma (TRG, formerly known as TCRG) chain studies involving: locus spans 160 kilobases on chromosome 7 (7p14). The locus consists of 14 variable (V) gene segments in six subgroups, and five joining Identification of clonal T-cell populations highly suggestive of (J) gene segments interspersed between two constant (C) geneT-cell malignancies segments. However, the repertoire of functional TRG molecules Lineage determination of leukemias and lymphomas is limited to 4-6 functional V gene segments that belong to two subgroups. 2Monitoring and evaluation of disease recurrenceDetection and assessment of residual disease Rearrangement of the V and J gene segments of the TRG locus results in V-Jproducts of unique length and sequence. Clonal TRG Evaluation of new research and methods in malignancy studies rearrangements can be most rapidly identified by analyzing the size distribution of DNA products amplified from conserved sequences that Summary and Explanation of the Testflank this V-Jregion. 1DNA isolated from a normal heterogeneous population of polyclonal T-cells produces a Gaussian distribution Sample genomic DNA is amplified using two master mixes that(bell-shaped size curve) of amplified products. DNA amplified from independently target conserved regions within the variable (V) anda clonal T-cell population generates one or two product(s) of unique joining (J) regions that flank the unique, hypervariable, antigen- size that reflects proliferation of a single rearranged clone. 1,2binding, complementarity determining region 3 (CDR3). This assay targets V1-9 and J gene segments. Positive and negative DNA controls, as well as an internal Amplification Control Master Mix, areSpecimen Requirementsincluded. The limit of detection of this assay is approximately one5 mL of peripheral blood, bone marrow biopsy, or bone marrow clonal T cell in a background of a hundred normal cells. PCR products can be analyzed by capillary electrophoresis or standard aspirate anti-coagulated with heparin or EDTA; or,gel electrophoresis with ethidium bromide staining. Minimum 5 mm cube of tissue; or,2 g of genomic DNA; or,Formalin-fixed, paraffin-embedded (FFPE) tissue or slides.Reference1.JE Miller, SS Wilson, DL Jaye, and M Kronenberg. J. Mol. Diag. 4:101-117 (1999).2. K Beldjord et al., Leukemia 17: 2289-2292 (2003). Figure Legend: Simplified figure representing the organization of a rearranged T-cell receptor gamma chain gene on chromosome 7. Colored arrows represent conserved regions within the variable region gene segments targeted by primers. Primers are represented by arrows with the size range of valid products generated with each of the master mixes indicated below the figure. Colors correspond to the peak colors assigned to products when differential fluorescence detection methods are used.112'