⚡ Detection Method

qPCR

🕒 Assay Time

Refer to manual

🎯 Storage Temperature

-20 °C (stable for 24 months)

Product Description

SHENTEK® CAR/TCR Transgene Copy Number Quantitation Kit is designed to quantify the copy number of target genes in human cell products derived from common HIV-1 lentiviral vectors (e.g., CAR or TCR genes integrated in CAR-T and TCR-T cell genomes). The kit uses a multiplex qPCR assay to detect DNA sequences related to the integration/expression elements on the transfer plasmid and a Single Copy Gene (SCG) in human cells, enabling calculation of average copies of target gene per cell. The kit includes a company-produced reference standard and an internal positive control (IPC). The assay is intended for rapid, specific, reliable and universal lentiviral vector applications. For Research Use Only.

Technical Specifications

Parameter	Details
Product number	SK030221CA100
Intended reactions	Reagents for 100 reactions
Reference standard initial concentration	CAR/TCR Control initial copy number: 2.33 × 10^8 copies/μL
Standard curve serial dilutions (copies/μL)	ST1: 2.33 × 10^6; ST2: 2.33 × 10^5; ST3: 2.33 × 10^4; ST4: 2.33 × 10^3; ST5: 2.33 × 10^2
Reaction volume	Total reaction volume 30 μL (20 μL qPCR MIX + 10 μL template)
qPCR MIX composition (per reaction)	qPCR Reaction Buffer 15.9 μL; CAR/TCR Primer & Probe MIX 2.8 μL; IPC MIX 1.3 μL (total 20 μL)
Thermal cycling program	Activation 95 °C 10:00 (1 cycle); Denaturation 95 °C 00:15 (40 cycles); Annealing/Extension 60 °C 01:00 (fluorescence read)
Reporter dyes / targets	CAR/TCR target: FAM; SCG: CY5; IPC: VIC; ROX as passive reference
Ct / analysis settings	Manual Ct threshold 0.02; Automatic baseline; verify slope/intercept/R² for standard curve
Acceptance criteria for IPC	Mean Ct-IPC of sample should be within ±1.0 of the NCS Ct-IPC; significant increase indicates inhibition
Recommended DNA input per well	Recommended 5 ng to 30 ng total DNA per well (adjust based on preliminary experiments)
Calculation formula	CAR or TCR copies/cell = 2 × (CAR or TCR / SCG)
Sample types	Purified genomic DNA from CAR-T, TCR-T or other cells with lentiviral integration

Features

Multiplex absolute quantitation: Simultaneous detection of CAR/TCR target, single copy gene (SCG) and IPC in the same reaction for absolute copy number determination using a standard curve.
Reference standard included: Proprietary reference standard supplied and pre-dilution scheme provided for a 5-point standard curve.
Ready-to-use primer & probe mixes: Pre-formulated CAR/TCR Primer & Probe MIX and IPC MIX simplify assay setup and reduce pipetting error.
Compatible with common qPCR platforms: Validated for use on SHENTEK-96S, ABI 7500, LineGene 9600plus, CFX96 and similar real-time PCR instruments.
Designed for lentiviral vector applications: Optimized for quantifying transgene copy numbers from HIV-1 derived lentiviral vectors (CAR/TCR).

Applications

Quantification of CAR or TCR transgene copy number in CAR-T and TCR-T cell products
Lentiviral vector integration assessment
Preclinical and research cell therapy product characterization
Quality control and batch testing for gene-modified cell products (research use)

Kit Contents

qPCR Reaction Buffer (NNB001) 850 μL × 2 tubes-20 °C, protect from light

CAR/TCR Primer & Probe MIX (NNC032) 300 μL × 1 tube-20 °C

IPC MIX (NNC066) 150 μL × 1 tube-20 °C

DNA Dilution Buffer (DDB) (NND001) 1.5 mL × 3 tubes2–8 °C (store remaining DDB at 2–8 °C); -20 °C for long term per kit table

CAR/TCR Control (NNA022) 50 μL × 1 tube-20 °C

* Total: 100 reactions | Method: qPCR (multiplex absolute quantitation, standard curve)

Attention

• For Research Use Only (RUO). Not for use in diagnostic procedures.

• Read Material Safety Data Sheets (MSDS) and follow handling instructions; wear protective eyewear, mask, clothing and gloves.

• Work in a clean environment: irradiate bench, pipettes and tubes with UV for 30 minutes and disinfect with 75% ethanol to avoid contamination.

• Avoid assay inhibitors; monitor IPC Ct — if sample IPC Ct is significantly higher than NCS IPC Ct, sample may be inhibitory.

• NTC and NCS Ct should be larger than the mean Ct of the lowest standard concentration.

• Prepare and run triplicates for standards, NTC, NCS and test samples where possible.

• Recommended sample input per well: 5 ng–30 ng total DNA (optimize as needed).

• Store kit components as labeled (-20 °C) and protect from light; kit stable up to 24 months. If DDB is cloudy, heat to 37 °C until clear.

• Seal plates/tubes, mix and briefly centrifuge before running; follow instrument-specific setup and analysis instructions.

Quality Management & Certifications

Quality System

Download QMS

QMS (ISO, GMP)

Download Certificate

Quality Advantages

View Report

Quality Control Process

Download Process

📂 Technical Resources & Downloads

PDF Product Manual ↓ PDF MSDS / SDS ↓

Frequently Asked Questions (FAQ)

Q1: What sample type and amount should I use for the assay?

Use purified genomic DNA from CAR-T, TCR-T or other cells with lentiviral integration. The recommended total DNA per well is between 5 ng and 30 ng; adjust based on preliminary experiments.

Q2: How is transgene copies per cell calculated?

Calculate CAR or TCR copies per cell using the formula: CAR or TCR copies/cell = 2 × (CAR or TCR / SCG), where CAR/TCR and SCG quantities are measured from the standard curve (copies/μL).

Q3: What are the qPCR thermal cycling conditions?

Activation 95 °C for 10:00 (1 cycle); Denaturation 95 °C for 00:15 (40 cycles); Annealing/Extension 60 °C for 01:00 with fluorescence read during this step.

Q4: How should I interpret IPC and control results?

Mean Ct-IPC of sample should be within ±1.0 of the NCS Ct-IPC; a significantly higher IPC Ct indicates possible inhibition. NTC and NCS Ct values should be larger than the mean Ct of the lowest standard.

Research Use Only

✔

Research Use Only (RUO) – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.

✔

Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.

CAR/TCR Transgene Copy Number Quantitation Kit SHENTEK®