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T Cell Receptor Gamma Gene Rearrangement Assay MegaKit - ABI Fluorescence Detection
Product Name Cat. No. Size Price Qty.
T Cell Receptor Gamma Gene Rearrangement Assay MegaKit - ABI Fluorescence Detection 1-207-0071
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Product Uses
Historically, T Cell Receptor Gamma Chain Gene Rearrangement Assays have been used to:
  1. Identify clonality highly suggestive of T-cell and some immature B-cell malignancies
  2. Determine lineage involvement in mature lymphoproliferative disorders
  3. Monitor and evaluate disease recurrence
Overview
Invivocribe Technologies' assay are standardized PCR-based tests. Each test comes with a Standard Operating Procedure (SOP), an interpretation guide, master mixes, and controls. 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. Click to read more
Description
The T cell receptor gamma (TCRG) chain locus spans 160 kilobases on chromosome 7 (7p14). The locus consists of 14 variable (Vg) gene segments in 6 subgroups, and 5 joining (Jg) gene segments interspersed between 2 constant (Cg) gene segments. However, the repertoire of functional TCRG molecules is limited to 4-6 functional Vg gene segments that belong to 2 subgroups. Rearrangement of the variable (Vg) and joining (Jg) gene segments of the TCRG locus results in Vg-Jg products of unique length and sequence. Clonal TCRG rearrangements can be most rapidly identified by analyzing the size distribution of DNA products amplified from conserved sequences that flank this Vg-Jg region. DNA isolated from a normal heterogeneous population of polyclonal T cells produces a Gaussian distribution (bell-shaped size curve) of amplified products. DNA amplified from a clonal T cell population generates one or two product(s) of unique size that reflects proliferation of a single rearranged clone.

Sample genomic DNA is amplified using two master mixes that independently target conserved regions within the variable (Vg) and joining (Jg) regions that flank the unique, hypervariable, antigen-binding, complementarity determining region 3 (CDR3). This assay targets Vg1-9 and Jg gene segments. Positive and negative DNA controls, as well as an internal Amplification Control master mix, are included. Click to read more
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.
Materials Provided
QTY Description
10 Amplification Control Master Mix - 6FAM
10 T Cell Receptor Gamma Mix 1 - 6FAM
10 T Cell Receptor Gamma Mix 2 - HEX
5 IVS-0000 Polyclonal Control DNA
5 IVS-0009 Clonal Control DNA


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