b'Gel & Capillary | RUO AssaysGel and CapillaryIGL Gene Clonality AssaysResearch Use Only (RUO) AssayIGL Gene Clonality AssaysAssay Use IGL gene rearrangements (V-J) rearrangements potentially represent an attractive PCR target for clonality studies to compensate Immunoglobulin lambda light chain (IGL) gene clonality assays arefor false-negative IGH V H -J HPCR results mainly caused by somatic useful for the identification of B-cell clonality, studying clonal B-cellhypermutations. The limited size of the junctional region may createpopulations, and evaluation of new research and methods a challenge to distinguish polyclonal from monoclonal in malignancy studies. rearrangements when running a simple agarose or polyacrylamide gel. 1 Therefore, clonal V-J PCR products are most easily and reliably Summary and Explanation of the Testidentified by heteroduplex analysis using standard polyacrylamide gels. Alternatively, capillary electrophoresis or gene sequencing The IGL Tube master mix targets conserved regions within theinstruments coupled with differential fluorescence detection can also variable (V1-3) and the joining (J1-3) regions that flank the unique,be used for analysis. 1hypervariable, antigen-binding, complementarity determining region 3 (CDR3). Positive and negative controls, as well as a Specimen Control Size Ladder Master Mix, are included. PCR products can be analyzed by capillary electrophoresis or heteroduplex analysis.Specimen RequirementsClonality is indicated if the master mix generates clonal products. This assay tests extracted and purified genomic DNA (gDNA).BackgroundReferencesThe human immunoglobulin lambda (IGL) light chain locus is1.F Davi et al., Leukemia 17:2280-2283 (2003).located on the long arm of chromosome 22 (22q11.2) and spans2. JJM van Dongen et al., Leukemia 17:2257-2317 (2003).1050 kilobases. It is made up of 73-74 variable (V) gene segments (spread over 900 kilobases), 7-11 joining (J) gene segments and7-11 constant (C) gene segments depending on the haplotypes.Of the 73-74 V region genes, only 30-33 are functional and can be 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.106'