Phusion™ Plus Green PCR Master Mix (Thermo Scientific™)

Thermo Scientific Phusion Plus Green PCR Master Mix is a convenient, ready-to-use 2X mixture of Phusion Plus DNA Polymerase, buffer, magnesium chloride, and dNTPs. The master mix format simplifies reaction setup with minimal pipetting steps and retains all features of Phusion Plus DNA Polymerase. Additionally, two tracking dyes and a density reagent are pre-added to the master mix for direct loading of PCR products on gels.

• 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

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 Green PCR Master Mix to generate PCR sequences with high accuracy, sensitivity, and inhibitor tolerance. Its novel 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 Green PCR Master Mix and its universal annealing feature, different PCR reactions can be cycled together using the same protocol.

Phusion Plus Green PCR Master Mix 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
• Amplification of difficult (GC-rich) templates
• Long-range PCR (up to 20 kb)
• Mutagenesis
• High-throughput PCR

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