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For Research Use Only (RUO)
$1,545.00
Sku: 1101111-1
Product Description
SHENTEK® Residual Plasmid DNA Quantitation Kit (3G) is a duplex real-time PCR (qPCR) kit designed to quantify residual plasmid DNA in gene therapy products (for example, plasmid DNA in lentiviral vector preparations used in CAR‑T manufacturing). The assay targets consensual plasmid sequences (replicon sequences of ColE1/pMB1/pBR322/pUC) and uses a FAM-labeled target probe for quantitative detection and a VIC-labeled Internal Positive Control (IPC) included in the Primer&Probe MIX to monitor reaction performance. The kit provides reagents and a lyophilized plasmid linear DNA control for generation of a standard curve, enabling reliable quantitation down to the 10^2 copies/µL level. The kit is supplied for 100 reactions and is intended for research use only. Compatible with common real-time PCR platforms (e.g., SHENTEK-96S, ABI 7500, Bio‑Rad CFX96, Roche LightCycler 480).
Technical Specifications
| Parameter | Details |
|---|---|
| Limit of detection (LOD) | Approximately 1×10^2 copies/µL (sensitivity reported by manufacturer) |
| Quantitative dynamic range / Standard curve | Serial dilution standard curve covers 4.97×10^8 to 4.97×10^1 copies/µL (prepared from stock A down through ST5) |
| Reaction volume | 30 µL total (20 µL qPCR MIX + 10 µL sample/standard/control) |
| qPCR mix per reaction | qPCR Master MIX 15 µL + Plasmid Primer&Probe MIX (incl. IPC) 5 µL = 20 µL qPCR MIX |
| Thermal cycling conditions | Activation: 95 °C 10:00 (1 cycle); 40 cycles of 95 °C 00:15 (denaturation) and 60 °C 01:00 (anneal/extend; fluorescence read during this step) |
| Detection channels / probes | Target probe: FAM (Plasmid-DNA); Internal Positive Control (IPC): VIC |
| Threshold / analysis settings (recommended) | Manual Ct threshold: Plasmid-DNA (FAM) = 0.05; IPC (VIC) = 0.1. Automatic baseline recommended. |
| Quality acceptance criteria | ERC recovery rate acceptance: 50%–150%; NTC Ct should be ≥ (ST5 Ct + 2) or lab-specific; NCS Ct should be larger than mean Ct of lowest standard and show normal VIC amplification |
| Storage & shelf life | -20 °C (components stable for up to 24 months under recommended storage) |
| Kit capacity | Reagents for 100 reactions |
Features
- Duplex qPCR with IPC: Includes an Internal Positive Control (VIC) within the Primer&Probe MIX to monitor PCR performance and detect inhibition or reaction failure.
- High sensitivity: Reported detection sensitivity at the 10^2 copies/µL level for trace plasmid DNA quantitation.
- Consensus plasmid sequence targeting: Assay targets consensual replicon sequences (ColE1/pMB1/pBR322/pUC) to detect a broad range of common plasmid backbones.
- Ready-to-use reagents and controls: Includes lyophilized plasmid linear DNA control and pre-mixed Primer&Probe and Master MIX reagents to streamline setup.
- Broad instrument compatibility: Validated for use on common real-time PCR systems (SHENTEK-96S, ABI 7500, Bio‑Rad CFX96, Roche LightCycler 480 and similar platforms).
- Workflow for regulated testing environments: Provides recommended standard curve design, plate layouts, and acceptance criteria to support method validation and sample testing.
Applications
- Quantitation of residual plasmid DNA in gene therapy products
- Monitoring plasmid DNA residue in lentiviral vector preparations (CAR‑T production workflows)
- Quality control and release testing for biologics where plasmid backbone contamination must be measured
- Method validation and process development for plasmid removal/clearance
Kit Contents
Plasmid linear DNA Control (NNA016), lyophilized powder ×1 tube-20 °C
Plasmid Primer&Probe MIX (Incl. IPC) (NNC120), 500 µL ×1 tube-20 °C, protect from light
qPCR Master MIX (NNB023), 850 µL ×2 tubes-20 °C, protect from light
DNA Dilution Buffer (DDB) (NND001), 1.5 mL ×3 tubes-20 °C
* Total: 100 reactions | Method: qPCR (duplex real-time PCR)
Attention
• For research use only (not for diagnostic or clinical use).
• Read Material Safety Data Sheets (MSDS) and observe all handling instructions.
• Wear appropriate PPE (protective eyewear, mask, clothing and gloves) when handling reagents and samples.
• Use DNase-free, low-retention tubes and tips to avoid contamination and sample loss.
• Decontaminate work surfaces, pipettes and tubes with UV (30 minutes) and 75% ethanol before experiments to minimize nucleic acid contamination.
• Thaw reagents on ice or at 2–8 °C, vortex and briefly centrifuge before use. Protect light-sensitive reagents from light.
• Store all kit components at -20 °C. Unused DNA Dilution Buffer (DDB) should be stored at 2–8 °C; clear cloudy DDB by warming to 37 °C if necessary.
• Include recommended controls (NTC, NCS, ERC, and standard curve points) and run samples in triplicate when possible.
• Follow the provided plate layouts, thermal profile and analysis settings; adjust thresholds/criteria only after laboratory validation.
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What is the assay sensitivity (limit of detection)?
The kit is reported to detect residual plasmid DNA down to approximately 1×10^2 copies/µL. Actual limit of detection should be confirmed by the user's in-house validation.
Q2: How should I prepare the standard curve?
Dissolve the lyophilized Plasmid linear DNA Control in 55 µL ddH2O, dilute to the stock concentration indicated on the tube, then perform serial 1:10 dilutions to generate standards A → ST5. Recommended standard points in the manual range from 4.97×10^8 to 4.97×10^1 copies/µL.
Q3: How do I interpret ERC recovery and control criteria?
ERC (extraction reference control) recovery rates should fall between 50% and 150%. NCS Ct should be larger than the mean Ct of the lowest standard and show VIC amplification; NTC Ct should be ≥ ST5 Ct + 2 (or per your lab validation).
Q4: How should the kit be stored and what is the shelf life?
Store all kit components at -20 °C and protect light-sensitive reagents from light. Components are stable for up to 24 months when stored under recommended conditions.
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.
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
