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

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

Catalog No. F534S
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Catalog No. Color No. of Reactions
F534S Green 100 Reactions
F530S Colorless 100 Reactions
F530L Colorless 500 Reactions
F530XL Colorless 2000 Reactions
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Catalog No. F534S Supplier Thermo Scientific™ Supplier No. F534S
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Description

Includes

Phusion High-Fidelity DNA Polymerase

  • Phusion DNA Polymerase 2 U/μL
  • 5X Phusion HF Buffer
  • 5X Phusion GC Buffer
  • DMSO
  • 50 mM MgCl2 solution

Both Phusion HF Buffer and Phusion GC Buffer provide 1.5 mM MgCl2 in the final 1X concentration.

Phusion Green High-Fidelity DNA Polymerase

  • Phusion DNA Polymerase 2 U/μL
  • 5X Phusion Green HF Buffer
  • 5X Phusion Green GC Buffer
  • DMSO
  • 50 mM MgCl2 solution

Both Phusion Green HF Buffer and Phusion Green GC Buffer provide 1.5 mM MgCl2 in the final 1X concentration.

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 Green HF and GF Buffers
• DMSO
• 50 mM MgCl2

Phusion Green HF and Phusion Green 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 Green
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Frequently Asked Questions (FAQs)

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.
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.
Can I use Phusion Green and Phire Green PCR products for subsequent applications such as sequencing, ligation, and restriction digestion?

Yes. The dyes do not interfere with downstream applications such as DNA sequencing, ligation, and restriction digestion.
What dyes are present in Phusion Green and Phire Green buffers?

Phusion Green and Phire Green buffers contain two dyes for monitoring electrophoresis progress. The blue dye migrates with 3-5 kb DNA fragments and the yellow dye migrates faster than 10 bp DNA fragments in 1% agarose gel. The dyes have excitation peaks at 424 nm and 615 nm, respectively.

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