b'Gel & Capillary | RUO AssaysGel and CapillaryIGK Gene Clonality AssaysResearch Use Only (RUO) AssayIGK Gene Clonality AssaysAssay UseIGK Gene Clonality Assays are useful for studies involving: encode the first 95 N-terminal amino acids. Positions 96-108 are encoded by one of five joining (J) gene segments. The constant (C)dentification of clonal B-cell populations highly suggestive ofI portion of the kappa light chain (amino acids 109-214) is encoded by a B-cell malignancies single constant (C) region separated from the J region by an intron. Lineage determination of leukemias and lymphomas The length of the hypervariable complementarity determining region 3 (CDR3) in kappa light chain genes is limited and rearrangements in Monitoring and evaluation of disease recurrence this region display significant skewing (platykurtosis). 2Detection and assessment of residual diseaseEvaluation of new research and methods in malignancy studies Therefore, clonal CDR3 products generated from this region are most easily and reliably identified by heteroduplex analysis using standard polyacrylamide gels. Alternatively, capillary electrophoresis Summary and Explanation of the Testor gene sequencing instruments coupled with differential fluorescence detection can be used for analysis.Two master mixes target conserved regions within the variable(V1-7) and the joining (J1-5) regions that flank the unique hypervariable, antigen-binding, complementarity determiningSpecimen Requirementsregion 3 (CDR3). Other primers target the K deand intragenic regions. Tube A contains six upstream primers and two J region primers. 5 mL of peripheral blood, bone marrow biopsy, or bone marrow Tube B contains six upstream V region primers, an upstreamaspirate anti-coagulated with heparin or EDTA; or,intragenic primer and a downstream K deprimer. Positive andMinimum 5 mm cube of tissue; or,negative controls, as well as a Specimen Control Size Ladder Master2 g of genomic DNA; or,Mix, are included. PCR products can be analyzed by capillary electrophoresis or heteroduplex analysis. Clonality is indicated if anyFormalin-fixed, paraffin-embedded (FFPE) tissue or slides.one of the master mixes generates clonal products.ReferenceBackground1.AW Langerak et al., Leukemia 17: 2275-2280 (2003).The human immunoglobulin kappa (IGK) light chain locus on the2. EP Rock, PR Sibbald, MM Davis, and YH Chien. J. Exp. Med. 179(1):short arm of chromosome 2 (2p12, formerly 2p11.2) spans 1820 kb. It is323-328 (1994).made up of 76 variable (V) gene segments belonging to 7 subgroups,3. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003).5 joining (J) gene segments, and one constant (C) gene segment. Productive assembly of the kappa gene is successful in about 60% of human B lymphocytes. 1However, even when unsuccessful, clonal B cells generally retain the rearranged kappa genes. The V segments This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Schematic diagram of the immunoglobulin kappa light chain gene complex on chromosome 2p11.2. Shown are the relative positions and orientations for the V-J, and K deprimers, which are included in the IGK master mix tubes.102'