b'Gel & Capillary | RUO AssaysGel and CapillaryT-Cell Receptor Gamma Gene Research Use Only (RUO) AssayRearrangement AssaysT-Cell Receptor Gamma Gene Rearrangement AssaysAssay Use Specimen RequirementsT-Cell receptor gamma (TCRG)gene clonality assays are useful forThis assay tests extracted and purified genomic DNA (gDNA).the identification of T-cell clonality, studying clonal T-cell populations, and evaluation of new research and methods in malignancy studies. ReferencesSummary and Explanation of the Test1.JE Miller, SS Wilson, DL Jaye, and M Kronenberg. J. Mol. Diag. 4:101-117 (1999).Sample genomic DNA is amplified using two master mixes that2. K Beldjord et al., Leukemia 17: 2289-2292 (2003). independently target conserved regions within the variable (V) and joining (J) regions that flank the unique, hypervariable, antigen-binding, 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, are included. PCR products can be analyzed by capillary electrophoresis or standard gel electrophoresis with ethidium bromide staining.Background The T-cell receptor gamma (TRG, formerly known as TCRG) chain locus spans 160 kilobases on chromosome 7 (7p14). The locus consists of 14 variable (V) gene segments in six subgroups, and five joining (J) gene segments interspersed between two constant (C) gene segments. However, the repertoire of functional TRG molecules is limited to 4-6 functional V gene segments that belong to two subgroups. 2Rearrangement of the V and J gene segments of the TRG locus results in V-Jproducts of unique length and sequence. Clonal TRG rearrangements can be most rapidly identified by analyzing the size distribution of DNA products amplified from conserved sequences that flank this V-Jregion. 1DNA isolated from a normal heterogeneous population of polyclonal T-cells produces a Gaussian distribution (bell-shaped size curve) of amplified products. DNA amplified from a clonal T-cell population generates one or two product(s) of unique size that reflects proliferation of a single rearranged clone. 1,2Figure 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'