Thermo Scientific™ Phusion Hot Start II High-Fidelity PCR Master Mix

The Thermo Scientific™ Phusion Hot Start II High-Fidelity PCR Master Mix combines ultra-high fidelity DNA amplification with hot-start activation to deliver robust, specific, and high-yield PCR performance. This master mix features the Phusion Hot Start II DNA Polymerase, which provides 52X higher fidelity compared to Taq polymerase and eliminates non-specific amplification during setup.

The hot-start functionality is achieved via a proprietary Affibody® ligand, which inhibits polymerase activity at room temperature, including its 3’→5’ exonuclease function, to protect primers and templates from degradation. Upon thermal cycling, the ligand dissociates rapidly, ensuring zero activation time and immediate enzyme functionality.

The Green Master Mix variant includes tracking dyes and a density reagent for direct gel loading, while the Colorless Master Mix is ideal for fluorescent detection systems or downstream enzymatic applications.

Key Features

• Ultra-high fidelity: 52X Taq accuracy for error-sensitive applications
• Built-in hot-start via reversible Affibody® ligand – no activation step needed
• Fast PCR: Short extension times (15–30 s/kb)
• Reduced primer degradation during setup
• Ready-to-use 2X formulation with MgCl₂ and dNTPs
• Green format includes loading dyes for gel electrophoresis
• High specificity and sensitivity for complex or GC-rich templates
• Blunt-end PCR products ideal for cloning

Applications

• High-fidelity PCR and hot-start PCR
• Long-range PCR (up to 20 kb)
• Amplification of GC-rich or complex templates
• DNA template preparation for sequencing or cloning
• Multiplex PCR
• Mutagenesis workflows
• High-throughput PCR
• Microarray target generation

Catalog Numbers & Formats:

Concentration: 2X Master Mix

Storage: –20°C

Shipping Condition: Dry Ice

For Research Use Only

Kit Contents

• 2X Phusion Hot Start II High-Fidelity PCR Master Mix
• Nuclease-free water
• DMSO (optional for GC-rich templates)

Final 1X buffer contains 1.5 mM MgCl₂.

Specifications

The Phusion Hot Start II High-Fidelity PCR Master Mix offers a superior blend of speed, specificity, and accuracy for demanding PCR workflows. Whether you're cloning, sequencing, or amplifying difficult templates, this master mix provides robust performance with minimal optimization—ideal for both routine and high-throughput research applications.