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

Catalog No.	Color	No. of Reactions
F530S	Colorless	100 Reactions
F530L	Colorless	500 Reactions
F530XL	Colorless	2000 Reactions
F534S	Green	100 Reactions
F534L	Green	500 Reactions

Catalog No. F530S Supplier Thermo Scientific™ Supplier No. F530S

Description

Thermo Scientific Phusion High-Fidelity DNA polymerases set a gold standard for high performance PCR.

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.

Specifications

• Concentration: 2 U/μL
• Content And 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.
• GC-Rich PCR Performance: High
• Polymerase: Phusion High-Fidelity DNA Polymerase
• Reaction Speed: Fast
• Product Type: High-Fidelity DNA Polymerase
• Quantity: 100 units
• Shipping Condition: Dry Ice
• For Use With (Application): Standard PCR, High-fidelity PCR
• Fidelity (vs. Taq): 52X
• Hot Start: No
• No. of Reactions: 100 Reactions
• Overhang: Blunt
• Reaction Format: Standalone
• Size (Final Product): 20 kb or less
• Color: Colorless

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.

What nucleotide analogues can I use with DyNAzyme and Phusion DNA Polymerases?

DyNAzyme II DNA Polymerase can use dUTP, biotinylated dNTPs, 7-deaza-dGTP, digoxigenin-dUTP, bromo-dUTP, radiolabeled dNTPs and ITP. DyNAzyme EXT DNA Polymerase and Phusion DNA Polymerase cannot read dUTP-derivatives or dITP in the template strand so the use of these analogues is not recommended. Use Phusion U Hot Start DNA Polymerase for amplification of dUTP and dITP containing templates.

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

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