b'Gel & Capillary | RUO AssaysGel and CapillaryIGL Gene Clonality AssaysResearch Use Only (RUO) AssayIGL Gene Clonality AssaysAssay UseIGL Gene Clonality Assays are useful for studies involving: IGL gene rearrangements are present in 5-10% of Ig kappa B-cell malignancies and in all Ig lambda B-cell malignancies. Therefore,Identification of clonal B-cell populations highly suggestiveV-J rearrangements potentially represent an attractive extra PCR of B-cell malignancies target for clonality studies to compensate for false-negative IGHLineage determination of leukemias and lymphomas V H -J HPCR results mainly caused by somatic hypermutations. It should Monitoring and evaluation of disease recurrence be noted that because of the limited size of the junctional region, it is extremely difficult to distinguish polyclonal from monoclonal Detection and assessment of residual disease rearrangements by running a simple agarose or polyacrylamide Evaluation of new research and methods in malignancy studies gel 1 . Therefore, clonal V-J PCR products are most easily and reliably identified by heteroduplex analysis using standard polyacrylamide gels. Alternatively, capillary electrophoresis or gene sequencing Summary and Explanation of the Testinstruments coupled with differential fluorescence detection can be used for analysis. 1The IGL Tube master mix targets conserved regions within the variable (V1-3) and the joining (J1-3) regions that flank the unique, hypervariable, antigen-binding, complementarity determining region 3 (CDR3). Positive and negative controls, as well as a SpecimenSpecimen RequirementsControl Size Ladder Master Mix, are included. PCR products canbe analyzed by capillary electrophoresis or heteroduplex analysis.5 mL of peripheral blood, bone marrow biopsy, or bone marrow Clonality is indicated if the master mix generates clonal products. aspirate anti-coagulated with heparin or EDTA; or,Minimum 5 mm cube of tissue; or,Background2 g of genomic DNA; or,The human immunoglobulin lambda (IGL) light chain locus isFormalin-fixed, paraffin-embedded (FFPE) tissue or slides.located on the long arm of chromosome 22 (22q11.2) and spans 1050 kilobases. It is made up of 73-74 variable (V) gene segmentsReference(spread over 900 kilobases), 7-11 joining (J) gene segments and7-11 constant (C) gene segments depending on the haplotypes. 1.F Davi et al., Leukemia 17:2280-2283 (2003).Of the 73-74 V region genes, only 30-33 are functional and can be2. JJM van Dongen et al., Leukemia 17:2257-2317 (2003).grouped into 11 families and 3 clans. 1The J and C region genes are organized in tandem with a J segment preceding a C gene. Typically there are 7 J-C segments of which four are functional and encode the four Ig lambda isotypes.This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.Figure Legend: Schematic diagram of the immunoglobulin lambda light chain gene complex on chromosome 22q11.2. Shown are the relative positions and orientations for the V and J primers, which are included in the IGL master mix tube. The two V primers only target V1, 2, and 3 because these three V families cover approximately 70% of rearrangeable V gene segments, and approximately 90% of all IGL gene rearrangements involve these three families. Similarly, the single J primer only targets J1, 2, and 3 because these three J segments are involved in 98% of all IGL gene rearrangements.104'