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CAT #: 23080010

BCL1/JH Tube - Unlabeled

Introduction

Master mixes are components of complete assays. Master mixes are composed of a buffered magnesium solution, deoxynucleotides, and multiple primers that target the gene segments of interest. Multiple primers are used to ensure a more comprehensive testing approach necessary to reliably identify clonal rearrangements. These tests are complete with the exception of Taq DNA Polymerase, which is not provided. A single thermocycler program and similar detection methods are used within each series of test to improve consistency, reduce human error, and facilitate cross training.

Standard Protocol:
50µl Total PCR Reaction Volume:

  1. Using gloved hands, remove the master mixes from the freezer. Allow the tubes to thaw; then gently vortex to mix.
  2. In a containment hood or dead air box remove an appropriate aliquot to clean, sterile microfuge tube (one tube for each of the master mixes). Aliquot volumes should be 45µl for each sample + 135µl for the positive, negative and no template controls. We recommend adding an additional 20µl to correct for pipetting errors.
  3. Add the appropriate amount of either AmpliTaq Gold or AmpliTaq DNA polymerase (0.25µl of either AmpliTaq Gold or AmpliTaq @ 5U/µl per 50µl total PCR reaction volume) to each of the master mixes and gently mix by inverting several times or gently vortexing.
  4. Aliquot 45µl of master mix to individual wells of a PCR plate.
  5. Add 5µl of DNA from the unknown and control samples to individual tubes or wells containing the respective master mix reactions, and pipette up and down several times to mix. Amplify target DNA using the universal thermocycler program.

Product Use

Historically, BCL1/JH t(11;14) Translocation Assays have been used to:

  1. Identify BCL1/JH gene rearrangements highly suggestive of mantle cell lymphoma
  2. Distinguish mantle cell lymphoma from other neoplastic or benign B cell proliferations
  3. Monitor and evaluate disease recurrence and detect residual disease

Product Details

  • Specimen Requirements

    This product tests genomic DNA. DNA can be extracted from the following specimens:

    1. 5cc of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA; OR,
    2. Minimum 5mm cube of tissue shipped frozen, at room temperature, or on ice in RPMI 1640; OR,
    3. Formalin-fixed paraffin embedded tissue or slides.
  • Description

    The BCL1/JH 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 Ig 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 BCL1/MTC locus can be detected by PCR methodology. However, breakpoints outside of the BCL1/MTC locus will not be identified by this particular test. Therefore, a negative result does not completely exclude the presence of a BCL1/JH gene rearrangement in the sample. The performance characteristics of this product have been independently determined by the EuroClonality Group.

    Two master mixes are included in this assay kit. The BCL1/JH master mix targets the major translocation cluster (MTC) of the BCL1 locus and the joining region of the Ig heavy chain locus. The Specimen Control Size Ladder master mix targets multiple genes and generates a series of amplicons of 100, 200, 300, 400, and 600 basepairs to ensure that the quality and quantity of input DNA is adequate to yield a valid result. Positive and negative controls are included. Clonality is indicated if the master mix generates clonal band(s). PCR products can be analyzed using standard gel electrophoresis with ethidium staining.

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