Phusion™ High-Fidelity DNA Polymerases (2 U/μL)
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Phusion™ High-Fidelity DNA Polymerases (2 U/μL)
Thermo Scientific™

Phusion™ High-Fidelity DNA Polymerases (2 U/μL)

Thermo Scientific Phusion High-Fidelity DNA polymerases set a gold standard for high performance PCR.
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Catalog NumberColorNo. of Reactions
F530SColorless100 Reactions
F530LColorless500 Reactions
F-530XLColorless2000 Reactions
F534SGreen100 Reactions
F534LGreen500 Reactions
Catalog number F530S
Price (USD)
137.00
Each
Add to cart
Color:
Colorless
No. of Reactions:
100 Reactions
Request bulk or custom format
Price (USD)
137.00
Each
Add to cart
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 an 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 polymerases.

Features of Phusion™ High-Fidelity DNA Polymerase include:

  • High fidelity (52X Taq)
  • Fast PCR due to short extension times (15–30 s/kb)
  • Robust performance, minimal optimization needed
  • High yields of PCR products with minimal enzyme amounts
  • Available in Green buffer format for direct loading of PCR products on gels (F-534S or F-534L)

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
  • Microarray

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

For Research Use Only. Not for use in diagnostic procedures.
Specifications
GC-Rich PCR PerformanceHigh
PolymerasePhusion High-Fidelity DNA Polymerase
Reaction SpeedFast
Product TypeHigh-Fidelity DNA Polymerase
Quantity100 U
Shipping ConditionDry Ice
For Use With (Application)Standard PCR, High-fidelity PCR
Concentration2 U/μL
Fidelity (vs. Taq)52X
Hot StartNo
No. of Reactions100 Reactions
OverhangBlunt
Reaction FormatStandalone
Size (Final Product)20 kb or less
ColorColorless
Unit SizeEach
Contents & Storage
• Phusion DNA Polymerase (2 U/μL)
• 5X Phusion HF & GC Buffers
• DMSO
• 50 mM MgCl2 solution

Phusion HF Buffer and Phusion GC Buffers each provide 1.5 mM MgCl2 in the final 1X concentration.
Store at -20°C.

Frequently asked questions (FAQs)

What is enzyme concentration in Phusion Hot Start II High-Fidelity PCR Master Mix?

Phusion Hot Start II DNA polymerase concentration is optimized to give good results in most reactions. When the PCR reaction is set up according to the instructions, the final concentration of Phusion enzyme is 1 U in 50 µL reaction (0.4 U in 20 µL reaction).

Do Phusion DNA Polymerases add the non-template dependent 3'-A overhang?

Phusion DNA Polymerases generate blunt end products; therefore, blunt end cloning is recommended. If TA cloning is required, it can be performed by adding A overhangs to the blunt PCR product with e.g. Taq DNA Polymerase (Cat. No. EP0401). However, before adding the overhangs it is very important to remove all the Phusion DNA Polymerase by purifying the PCR product carefully, as the proofreading activity in Phusion DNA Polymerase is very strong. Any remaining Phusion DNA Polymerase will degrade the A overhangs, thus creating blunt ends again.

Can Phusion DNA Polymerases extend at 1 second/kb?

Yes it is possible, especially when amplifying smaller amplicons. Processivity of Phusion DNA Polymerases is 10 times that of Pfu. We recommend extension times of 15 s/kb for Phusion Flash PCR Master Mix. 15 s/kb is a conservative value that we can promise to work with almost any amplicon. In many cases, significantly shorter extension times (0-5 s/kb) can be used without compromising the yield. What separates Phusion Flash DNA Polymerase from other fast polymerases is that all steps in the PCR protocol can be shortened, including annealing and denaturation. This results in extremely fast protocols as compared with any other polymerase.

Can protocols optimized for Phusion DNA Polymerase be directly applied to Phusion Hot Start II DNA Polymerase?

Yes, protocols optimized for Phusion DNA Polymerase can be applied to Phusion Hot Start II DNA Polymerase reactions.

Do Phire and Phusion Hot Start II DNA Polymerases need a separate activation step in the PCR protocol?

No separate activation step is required since Phire and Phusion Hot Start II DNA Polymerases are inactivated by a reversibly bound, specific Affibody ligand that dissociates during initial denaturation.

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