Phusion Hot Start II High-Fidelity DNA Polymerase

  • Reaction set up at room temperature
  • Zero-time reactivation (Hot start)
  • 52x higher fidelity vs Taq
  • Short extension times (15-30 s/kb)
  • Higher yields and lower enzyme amounts vs other DNA polymerase
  • Broad application possibilities

Hot Start PCR setup at room temperature

The Phusion Hot Start II High-Fidelity DNA Polymerase enzyme is inactive at room temperature, due to the binding of a specific Affibody ligand, which allows you to easily avoid the production of nonspecific products. The Affibody ligand also inhibits the 3´-5´ exonuclease activity of the polymerase, hereby preventing degradation of primers and template DNA during reaction set up.

Activated Phusion Hot Start II High-Fidelity DNA Polymerase

When polymerase activity temperature is reached, the Affibody ligand is released, hereby activating the enzyme. Phusion Hot Start II High-Fidelity DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols. The active enzyme generates blunt ends in the amplification products.

The high fidelity enzyme features an error rate 52-fold lower than that of Taq, which makes it an excellent choice for cloning and other applications requiring high fidelity. The enzyme is also available as a Green buffer format permitting direct loading on gels.

Broad application possibilities

You can use the Phusion Hot Start II High-Fidelity DNA Polymerase for a wide variety of applications:

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

Ready-to-use Hot Start PCR master mixes

For easy use, ready-to-use Hot Start PCR Master MIxes including the Phusion Hot Start II High-Fidelity DNA Polymerase are also available. Beside the enzyme, the 2X Master mixes include nucleotides and optimized reaction buffer including MgCl2. These solutions will save you pipetting time.

For even more convenience, you can use the Phusion Green HS II PCR Master Mix. This mix also includes two tracking dyes and a density reagent that allows for direct loading of PCR products on a gel. The dyes in the master mix do not interfere with PCR performance.

Ordering information

Westburg offers this product in The Netherlands, Belgium, Luxembourg,
If you are not located in these countries please visit the Thermo Scientific - Nucleic Acid Detection website to find your local distributor.
Cat no. Description
TH F-537L Phusion Green Hot Start II High-Fidelity DNA Polymerase - 500 U
TH F-537S Phusion Green Hot Start II High-Fidelity DNA Polymerase - 100 U
TH F-549L Phusion Hot Start II High-Fidelity DNA Polymerase - 500 U
TH F-549S Phusion Hot Start II High-Fidelity DNA Polymerase - 100 U
TH F-565L Phusion HSII HF PCR Master Mix 500 x 50 µL rxns
TH F-565S Phusion HSII HF PCR Master Mix 100 x 50 µL rxns
TH F-566L Phusion Green HSII PCR Master Mix 500 x 50 uL rxns
TH F-566S Phusion Green HSII PCR Master Mix 100 x 50 uL rxns