b'Gel & Capillary | RUO AssaysGel and CapillaryTCRD Gene Clonality AssaysResearch Use Only (RUO) AssayTCRD Gene Clonality AssaysAssay Use potential combinatorial diversity, the complementarity determining region 3 (CDR3) or junctional diversity is extensive due to the addition TCRD Gene Clonality Assays are useful for studies involving: of N regions, P regions, and random deletion of nucleotides bydentification of clonal T-cell populations highly suggestive of recombinases. This diversity is also extended by the recombination I of up to three D segments and therefore up to four N regions within T-cell malignancies the rearranged TRD locus. This limited germline diversity encoded at Lineage determination of leukemias and lymphomas the TRD locus in conjunction with extensive junctional diversity results Monitoring and evaluation of disease recurrence in a useful target for PCR analysis. TRD recombination events have been used most extensively as clonal markers in both T- and B-cell Detection and assessment of residual disease ALL. This standardized multiplex PCR assay detects the vast majority of clonal TRD gene rearrangements using a single multiplex master Evaluation of new research and methods in malignancy studies mix 1 . This assay provides rapid TCR clonality assessment, reducing or eliminating the number of Southern blot tests performed in the Summary and Explanation of the Testlaboratory. 2The detection rate of clonal TRD gene rearrangements The TCRD Tube Master Mix targets conserved regions withinusing this assay is exceptionally high. 1 the variable (V1-6), the diversity (D2-3) and the joining (J1-4) regions that flank the unique, hypervariable, antigen-binding, complementarity determining region 3 (CDR3) of the T-cell receptorSpecimen Requirementsdelta (TRD, formerly known as TCRD). Positive and negative5 mL of peripheral blood, bone marrow biopsy, or bone marrow controls, as well as a Specimen Control Size Ladder Master Mix, are included. PCR products can be analyzed by capillary electrophoresisaspirate anti-coagulated with heparin or EDTA; or,or heteroduplex analysis. Clonality is indicated if the master mixMinimum 5 mm cube of tissue; or,generates clonal products. 2 g of genomic DNA; or,Formalin-fixed, paraffin-embedded (FFPE) tissue or slides.Background The human T-cell receptor delta (TRD, formerly known as TCRD) geneReferencelocus is comprised of a cluster of 10 genes located on chromosome1.FL Lavender et al., Leukemia 17: 2292-2296 (2003).14 (14q11.2) spread over 60 kilobases, localized between the T-cell receptor alpha (TRA, formerly known as TCRA) variable (V) and2. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003).joining (J) gene segments. It is made up of eight variable (V), three diversity (D), and four joining (J) gene segments. 1At least five of the eight V gene segments can also rearrange to J gene segments and other V gene segments may also be utilized in TRD gene rearrangements in rare cases. Although the small number of V, D, and J gene segments available for recombination limits the This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Simplified diagram of a representative rearranged T-cell receptor delta gene showing the approximate placement of the upstream and downstream DNA primers. The numbers of primers and their specificity are listed for the TRD Tube Master Mix tube.116'