Larger pack sizes of Phusion Hot Start II DNA Polymerase with dNTPs now available.
Thermo Scientific™

Phusion Hot Start II DNA Polymerase (2 U/µL)

Catalog number:  F549S

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Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR. Featuring an error rate 50-fold lower than that of Taq, and 6-fold lower than that of Pfu, Phusion High-Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity. Phusion DNA Polymerases offer robust performance with short protocol times, even in the presence of PCR inhibitors, and generate higher yields with lower enzyme amounts than other DNA polymerase.

Using Phusion Hot Start II High-Fidelity DNA Polymerase, amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand that inhibits DNA polymerase activity at room temperature.
The Affibody ligand also inhibits the 3´→5´ exonuclease activity of the polymerase, thus preventing degradation of primers and template DNA during reaction set up. At temperatures that promote polymerase activity, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols.


• Reaction set up at room temperature
• No non-specific amplification and primer degradation during reaction set up
• Zero-time reactivation due to unique hot start technology
• High fidelity (52X Taq)
• Fast PCR due to short extension times (15-30 s/kb)
• Robust reactions, minimal optimization needed
• Increased product yields with minimal enzyme amounts
• Available as a Green buffer format permitting direct loading on gels (F-537S or F-537L)


• High-fidelity PCR
• High throughput
• Amplification of difficult (GC-rich) templates
• Template generation for sequencing
• Multiplex PCR
• Long-range PCR
• Cloning
• Mutagenesis
• Microarray

Using Phusion DNA Polymerases
Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases). For optimal results, use our Tm calculator at

For Research Use Only. Not for use in diagnostic procedures.


Hot Start: Built-In Hot Start
Product Line: Phusion
Quantity: 100U (2U/µL)
Fidelity (vs. Taq): 52 X
For Use With (Application): PCR Amplification
GC-Rich PCR Performance: High
Overhang: Blunt
Polymerase: Phusion Hot Start II DNA Polymerase
Product Type Specs: Hot Start DNA Polymerase
Purity: DHPLC
Reaction Format: Standalone
Reaction Speed: Fast

Contents & storage

• Phusion Hot Start II High-Fidelity DNA Polymerase (2 U/µL)
• 5X Phusion HF Buffer
• 5X Phusion GC Buffer
• 50 mM MgCl2 solution
Both Phusion HF and GC Buffers provide 1.5 mM MgCl2 in the final 1X concentration.Store at -20°C.