Phusion™ Plus DNA Polymerase (Thermo Scientific™)

Thermo Scientific Phusion Plus DNA Polymerase is a hot-start, high-fidelity DNA polymerase that brings together protein fusion technology and universal primer annealing. It is a Pyrococcus-like proofreading DNA polymerase fused to a processivity-enhancing domain, enabling Phusion Plus DNA Polymerase to generate PCR sequences with high accuracy, sensitivity, and inhibitor tolerance.

• High fidelity—provides >100X higher sequence accuracy than Taq polymerase
• Convenient setup—enables a universal annealing temperature of 60°C in PCR
• Enhanced specificity—hot-start mechanism reduces non-specific amplification and primer degradation
• Benchtop stability—automation enabled due to 24-hour stability of assembled reactions
• High inhibitor tolerance—works with DNA of suboptimal purity

The novel Phusion Plus reaction buffer includes proprietary additives that stabilize primer/template duplexes during the annealing step at 60°C. This unique buffer formulation increases PCR specificity without the need to optimize annealing temperature for each primer pair, helping save time and avoid mistakes in PCR runs. Furthermore, with Phusion Plus DNA Polymerase and its universal annealing feature, different PCR reactions can be cycled together using the same protocol.

Phusion Plus DNA Polymerase is inactive at ambient temperatures due to its hot-start mechanism mediated by Affibody molecules, allowing setup and storage of assembled PCR reactions at room temperature. Enzyme activity is fully restored after the initial denaturation step.

Applications
• High-fidelity PCR
• Cloning
• Template generation for sequencing
• GC-rich PCR
• Long-range PCR (up to 20 kb)
• Mutagenesis
• High-throughput PCR

To simplify PCR reaction setup, Phusion Plus DNA Polymerase is also available in ready-to-use master mix formats. Phusion Plus PCR Master Mix is a convenient 2X mix of enzyme, reaction buffer, magnesium chloride, and dNTPs. Only template and primers need to be added to the PCR reaction. Phusion Plus Green PCR Master Mix additionally contains a density reagent and two tracking dyes for direct loading of PCR products on gels.

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