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Thermo Scientific™ Phusion™ High-Fidelity PCR Master Mix (2X)
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Specifications:
| Application | PCR | ||
| Storage Temperature | -20°C | ||
| Product Type | PCR Master Mix | Forms | Liquid |
| Product Brand | Thermo Fisher Scientific™ | ||
| Product Grade | Molecular Biology | ||
Thermo Scientific™ Phusion™ High-Fidelity PCR Master Mix sets the gold standard for high-performance PCR, delivering exceptional accuracy, speed, and robustness. Built around Phusion High-Fidelity DNA Polymerase, this ready-to-use 2X master mix provides an error rate up to 50-fold lower than Taq and 6-fold lower than Pfu, making it ideal for applications where sequence accuracy is critical.
The master mix contains Phusion DNA Polymerase, dNTPs, MgCl₂, and optimized reaction buffer, significantly reducing pipetting steps and experimental variability. Two optimized buffer systems are available—HF Buffer for routine high-fidelity amplification and GC Buffer for challenging or GC-rich templates.
Key Features
- Ultra-High Fidelity – Up to 52× higher fidelity than Taq DNA polymerase
- Fast PCR Performance – Short extension times (15–30 s/kb)
- Convenient 2X Format – Reduced setup time and pipetting errors
- Robust Amplification – Performs well in the presence of inhibitors
- High Yield with Low Enzyme Usage
- Blunt-End PCR Products – Ideal for cloning and downstream processing
HF Buffer vs GC Buffer – Choosing the Right Master Mix
| Feature | HF Buffer | GC Buffer |
|---|---|---|
| Error Rate | 4.4 × 10⁻⁷ | 9.5 × 10⁻⁷ |
| Recommended Use | Default for high-fidelity PCR | GC-rich or complex templates |
| Template Complexity | Standard | GC-rich / secondary structures |
| Fidelity | Highest | High |
Recommendation: Use HF Buffer as the default master mix. Switch to GC Buffer when working with difficult or GC-rich templates.
Applications
- High-fidelity PCR
- Cloning
- Template generation for sequencing
- Long-range PCR (up to 20 kb)
- Amplification of GC-rich or difficult templates
- Site-directed mutagenesis
- High-throughput PCR workflows
Annealing Considerations
Annealing rules for Phusion DNA Polymerase differ from Taq-based systems. Optimal annealing temperatures are typically higher and should be calculated using primer Tm values specific to Phusion protocols.
Available Formats
| Catalog Number | Buffer Type | No. of Reactions | Contents |
|---|---|---|---|
| F531S | HF Buffer | 100 | 2X Phusion Master Mix with HF Buffer (2 × 1.25 mL) + 100% DMSO (1 × 500 µL) |
| F532S | GC Buffer | 100 | 2X Phusion Master Mix with GC Buffer (2 × 1.25 mL) + 100% DMSO (1 × 500 µL) |
| F531L | HF Buffer | 500 | 2X Phusion Master Mix with HF Buffer (10 × 1.25 mL) + 100% DMSO (2 × 500 µL) |
| F532L | GC Buffer | 500 | 2X Phusion Master Mix with GC Buffer (10 × 1.25 mL) + 100% DMSO (2 × 500 µL) |
Technical Specifications
| Parameter | Specification |
|---|---|
| Fidelity (vs Taq) | 52× |
| Hot Start | No |
| Reaction Speed | Fast |
| Maximum Amplicon Size | ≤ 20 kb |
| Concentration | 2X |
| GC-Rich PCR Performance | Medium |
| Reaction Type | Standard PCR, High-Fidelity PCR |
Thermo Scientific™ Phusion™ High-Fidelity PCR Master Mix is a trusted, high-performance solution for researchers requiring exceptional accuracy, speed, and reproducibility in PCR. With flexible HF and GC buffer formulations, robust inhibitor tolerance, and simplified workflow, it is an ideal choice for cloning, sequencing, mutagenesis, and long-range PCR applications.
- Buffer Type: With HF Buffer With GC Buffer
- Pack Size: 2 × 1.25 mL, Sufficient for 100 reactions 10 × 1.25 mL, Sufficient for 500 reactions