110 Gel & Capillary | RUO Assays Invivoscribe 2019 | 111 Gel & Capillary RUO Assays BCL1/JH Translocation Assay Assay Use The BCL1/JH Translocation Assay is useful for studies involving: • Identification of IGH-BCL1 (now known as IGH-CCND1) gene rearrangements highly suggestive of mantle cell lymphoma • Lineage determination of leukemias and lymphomas • Monitoring and evaluation of disease recurrence • Detection and assessment of residual disease • Evaluation of new research and methods in malignancy studies Summary and Explanation of the Test Two master mixes are included in this assay kit. The BCL1/JH Master Mix targets the major translocation cluster (MTC) of the CCND1 locus (formerly known as BCL1) and the joining region (JH) of the immunoglobulin heavy chain locus (IGH). The Specimen Control Size Ladder Master Mix targets multiple genes and generates a series of amplicons of approximately 100, 200, 300, 400, and 600 base pairs to ensure that the quality and quantity of input DNA is adequate to yield a valid result. Positive and negative controls are included. PCR products can be analyzed using standard gel electrophoresis with ethidium bromide staining. A CCND1 translocation is indicated if the master mix generates product(s) within the valid size range. Background The IGH-CCND1 t(11;14)(q13;q32) translocation is mainly found in mantle cell lymphomas, but has also been seen in B-prolymphocytic leukemia (10-20%), plasma cell leukemia, splenic lymphoma with villous lymphocytes, chronic lymphocytic leukemia (2-5%), and in multiple myeloma (20-25%). The t(11;14) brings about juxtaposition of the cyclin D1 gene with the immunoglobulin heavy chain gene. This leads to a marked increase in expression of cyclin D1 driven by the Ig heavy chain gene enhancer, located in the intron between the JH and constant region genes. The overexpression of cyclin D1 accelerates the passage of transformed cells through the G1 phase. Approximately 41% of the breakpoints on the CCND1/MTC locus can be detected by PCR methodology. However, breakpoints outside of the CCND1/MTC locus will not be identified by this particular test. Therefore, a negative result does not completely exclude the presence of an IGH-CCND1 gene translocation in the sample1 . The performance characteristics of this assay have been independently determined by the EuroClonality/ BIOMED-2 Group. This consortium of 47 molecular diagnostic laboratories tested hundreds of clinical samples, taking into account their lymphoproliferative status as defined by the WHO classifications2 . Specimen Requirements 1.  5 mL of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA; or, 2. Minimum 5 mm cube of tissue; or, 3. 2 μg of genomic DNA; or, 4. Formalin-fixed, paraffin-embedded (FFPE) tissue or slides. Reference 1. P Wijers et al., Leukemia 17: 2296-2298 (2003). 2. JJM van Dongen et al., Leukemia 17: 2257-2317 (2003). Reagents Ordering Information Catalog # Products Quantity 1-308-0010 BCL1/JH Translocation Assay - Gel Detection 33 reactions 1-308-0020 BCL1/JH Translocation Assay MegaKit - Gel Detection 330 reactions Agarose Gel Electrophoresis Detection The data shown was generated using the master mix indicated. Amplified products were run on a 2% agarose/TBE gel. Lane 1 is data generated testing the recommended 100% clonal control DNA; lane 2 is data generated testing a 10% dilution of the recommended clonal control DNA; lane 3 is data generated testing a 1% dilution of the recommended clonal control DNA; lane 4 is data generated testing a 0.1% dilution of the recommended clonal control DNA; and, lane 5 is data generated testing IVS-0000 Polyclonal Control DNA (Cat# 4-092-0010). This control is used as a negative control for many of our tests. A standard 100 base pair DNA size ladder was run in the lanes flanking the test samples. Controls Concentration Units in Assay Units in Assay MegaKit IVS-0010 Clonal Control DNA 200 μg/mL 1 x 100 μL tube 5 x 100 μL tubes IVS-0000 Polyclonal Control DNA 200 μg/mL 1 x 100 μL tube 5 x 100 μL tubes Master Mixes Target Units in Assay Units in Assay MegaKit BCL1/JH Tube MTC of CCND1 + IGH JH 1 x 1500 μL tube 10 x 1500 μL tubes Specimen Control Size Ladder Multiple Genes 1 x 1500 μL tube 10 x 1500 μL tubes Figure Legend: Schematic diagram of the IGH-CCND1 t(11;14) translocation showing the cyclin D1 (CCND1) gene on the left and the Ig heavy chain (IGH) gene on the right. Shown are the relative positions and orientations for the BCL1/MTC primer and the JH primer, which are included in the BCL1/JH Master Mix tube. This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.