b'Gel & Capillary | RUO AssaysGel and CapillaryT-Cell Receptor Gamma Gene Research Use Only (RUO) AssayRearrangement Assay 2.0 T-Cell Receptor Gamma Gene Rearrangement Assay 2.0Assay Use malignancies. This is critical for more comprehensive analysis of patient samples, as some T-cell lymphoproliferative disorders involve T-Cell Receptor Gamma Gene Rearrangement Assays are useful forV and J regions that would not be identified with a single V (1-8) and studies involving: J 1/J 2 primer set.Identification of clonal T-cell populations highly suggestive ofIn addition, the polyclonal background that results from the T-cell malignancies combination of all primers in a single tube produces a more robust Monitoring and evaluation of disease recurrence and easily interpreted signal with capillary electrophoresis, which aids Evaluation of new research and methods in malignancy studies in the interpretation of small peaks. Competitive amplification of all TRG gene rearrangements allows for identification of a quantitative threshold for a positive result and helps to avoid false positive results. Summary and Explanation of the TestThe average size of the TRG gene rearrangement PCR amplicons is 190 nucleotides, with a normal distribution of product sizes between This T-Cell Receptor Gamma Gene Rearrangement Assay 2.0159 and 207 nucleotides. This protocol should lead to improved represents an improved approach to PCR-based clonality testing ofproduct formation from formalin-fixed, paraffin-embedded (FFPE) lymphoproliferative disorders, as it can detect the vast majority ofsamples compared to other protocols that yield products of 260 TCR gamma gene rearrangements with a single multiplex masternucleotides or larger.mix. Importantly, this assay includes, in a single tube, primers for all known groups of TCR gamma variable (V) region genes and joining (J) region genes that are involved in rearrangements ofSpecimen RequirementsT-cell lymphomas. In addition, all reverse primers that target the J 5 mL of peripheral blood, bone marrow biopsy, or bone marrow region genes are conjugated with the 6FAM fluorophore. Positive and negative controls, as well as a Specimen Control Size Ladderaspirate anti-coagulated with heparin or EDTA; or,Master Mix are included. PCR products are analyzed by capillaryMinimum 5 mm cube of tissue; or,electrophoresis. 2 g of genomic DNA; or,BackgroundFormalin-fixed, paraffin-embedded tissue or slides.The human T-cell receptor gamma (TRG, formerly known as TCRG)Referencegene locus on chromosome 7 (7q14) includes 14 V genes belonging1.TC Greiner et al., JMD 4: 137-143 (2002).to four subgroups, five J segments, and two C genes spread over 200 kilobases. The diversity of this locus has historically complicated2. LC Lawnickie et al., JMD 5: 82-87 (2003).PCR-based testing. Our new multiplex PCR assay represents an3. Y Sandberg et al., Leukemia 21: 21 (2007).improvement over existing assays as it can detect the vast majority of TCR gamma gene rearrangements with a single multiplex master4. Armand, Marine et al. HemaSphere, 2019;3:3.mix. This master mix targets all conserved regions within the variable (V) and joining (J) region genes that are described in lymphoid This assay was developed by Invivoscribe.The performance of this assay was reviewed and validated by the EuroClonality/BIOMED-2 Group. 4Figure Legend: Simple representation of the organization of the T-cell receptor gamma gene on chromosome 7. Black arrows represent the relative positions of primers that target the variable region genes and the downstream joining region gene segments that are involved in rearrangements in T-cell lymphomas. The downstream primers are fluorescently labeled through the incorporation of a 6FAM fluorophore. The amplicon products generated from these rearrangements are detected by capillary electrophoresis.110'