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

InVivoScribe 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.
50ul 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 45ul for each sample + 135ul for the positive, negative and no template controls. We recommend adding an additional 20ul to correct for pipetting errors.
3. Add the appropriate amount of either AmpliTaq Gold or AmpliTaq DNA polymerase (0.25ul of either AmpliTaq Gold or AmpliTaq @ 5U/ul per 50ul total PCR reaction volume) to each of the master mixes and gently mix by inverting several times or gently vortexing.
4. Aliquot 45ul of master mix to individual wells of a PCR plate.
5. Add 5ul 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.

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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.

This product is covered by one or more of the following patents and patent applications owned by or exclusively licensed to Invivoscribe Technologies, Inc. (IVS):, United States Patent No. 7,785,783, United States Patent Application Number 10/531,106, European Patent Number EP 1549764B1 and other pending patent applications originating from European Patent Application Numbers 03756746.8 and 047326551.9 (16 countries), Japanese Patent Number JP04708029B2, Japanese Patent Application Number 2006-529437, Brazil Patent Application Number PI0410283.5, Canadian Patent Application Number 2525122, Indian Patent Application Number 5792/DELNP/2005, Mexican Patent Application Number PA/a/2005/012102, Chinese Patent Application Number 200480016603.5, and Korean Patent Application Number 10-2005-7021561.

Use of this product may require nucleic acid amplification methods such as Polymerase Chain Reaction (PCR). Any necessary license to practice amplification methods or to use amplification enzymes or equipment covered by third party patents is the responsibility of the user and no such license is granted by Invivoscribe Technologies, Inc., expressly or by implication. This product is sold FOR RESEARCH USE ONLY; not for use in diagnostic procedures.