T-Cell Receptor Gamma GeneRearrangement Assay 2.0
Now Available for Gel Detection
Assay Uses
This Research Use Only assay identifies T-cell receptor gamma (TCRG) chain gene rearrangements and is useful for theidentification of clonal T-cell populations and evaluation of new research and methods in malignancy studies.

Background
Invivoscribe’s T-Cell Receptor Gamma Gene Rearrangement Assay 2.0 represents an improved approach to PCR-based clonality
testing of T-cell lymphoproliferative disorders. This multiplex PCR assay can detect the vast majority of TCR gamma generearrangements with a single multiplex master mix that includes primers for all known groups of TCR gamma variable region
genes and joining region genes involved in rearrangements in T-cell lymphomas. Amplified products generated targeting theTCR gamma gene locus fall within a single size range to facilitate interpretation.

Specimen Requirements
This assay tests extracted and purified genomic DNA (gDNA).

Cell lines containing clonal T-cell rearrangements and one polyclonal (negative) sample were assessed using the T-Cell Receptor Gamma Gene Rearrangement
Assay 2.0 comparing detection methods. The data shown below was generated using 5% dilutions of DNA from each of the positive cell lines in polyclonal DNA orpolyclonal DNA alone. Following PCR amplification, the products were detected on an ABI 3500xL instrument (right) versus heteroduplexing then running on a PAGE
gel (left). The clonal products generated with the TCRG-6FAM master mix fall within a single contiguous size range of 159 bp to 207 bp (indicated by a red border):

5% IVS-0005 Clonal Control DNA
170 bp

5% IVS-0009 Clonal Control DNA
198 bp196 bp

5% IVS-016 Clonal Control DNA
167 bp

IVS-0000 Clonal Control DNA

191 bpM-0040 Rev 02 April 2021

Materials
The gel detection method requires additional reagents, not included in the kit:
Table 1. Materials Required (not provided)

Reagent/ Material

Recommended Reagents/ Materials & SuppliersPart Number

Ethidium Bromide

Thermo Fisher Scientific: UltraPureTM 10mg/mL Ethidium Bromide

15585-001

6% Polyacrylamide Gels

Thermo Fisher Scientific: Novex® TBE Gels (6%, 12 well)EC62652Box

TBE Running Buffer

Thermo Fisher Scientific: Novex® TBE Running Buffer (5X)

LC6675

Gel Loading Buffer

Thermo Fisher Scientific: 10X BlueJuiceTM Gel Loading Buffer,
Novex Hi- Density TBE Sample Buffer (5X)

10816-015, LC6678

100 bp DNA Ladder

Thermo Fisher Scientific: TrackItTM 100 bp DNA Ladder

10488-058

Gel Electrophoresis Unit

N/A

N/AMethod
The gel detection protocol is consistent with the standardized method in all Invivoscribe gel detection assays:
Gel Detection – Heteroduplex Analysis
1. Denature 20 μL of PCR products at 94°C for 5 minutes.
2. Re-anneal PCR products at 4°C for 60 minutes.
3. Assemble electrophoresis unit using a 6% non-denaturing polyacrylamide TBE gel (made with 1X TBE) and 0.5X TBE
running buffer.
4. Add 5 μL of ice-cold non-denaturing bromophenol blue loading buffer to samples.
5. Load 20 μL of mixture into wells of the gel.
6. Run gel at 110V for 2-3 hours or 40-50V overnight.
• Voltage and electrophoresis time depend on the PCR amplicon size, acrylamide gel thickness and type of PCR equipment.
• Voltage and run time can be adapted accordingly.
7. Stain the gels in 0.5 μg/mL ethidium bromide (in water or 0.5X TBE Buffer) for 5-10 minutes.
8. Destain the gels 2X in water for 5-10 minutes.
9. Visualize the gel using UV illumination.
10. Photograph the gel and interpret the data.

Ordering Information
Catalog #Products

Quantity

1-207-0101

T-Cell Gamma Receptor Gene Rearrangement Assay 2.0

33 Reactions

1-207-0111

T-Cell Gamma Receptor Gene Rearrangement Assay 2.0 Mega Kit

330 Reactions

These products are for Research Use Only (RUO). Not intended for diagnostic purposes.

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