Phusion® High-Fidelity DNA Polymerase

Catalog # Concentration Size List Price Quantity Your Price
M0530L 2000 units/ml 500 units $706.00
$635.40
M0530S 2000 units/ml 100 units $176.00
$158.40
Catalog # Size List Price Your Price
M0530L 500 units $706.00
$635.40
M0530S 100 units $176.00
$158.40
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

For high speed and high performance PCR. Manufactured and quality-controlled at New England Biolabs, Thermo Scientific® Phusion High-Fidelity DNA Polymerase offers both high fidelity and robust performance.

  • 50X higher fidelity than Taq
  • Robust reactions - maximal success with minimal optimization
  • Offered with multiple buffers for customized reaction setup for different DNA templates
High-Fidelity DNA Polymerases are important for applications in which the DNA sequence needs to be correct after amplification. Its unique structure, a novel Pyrococcus-like enzyme fused with a processivity-enhancing domain, increases fidelity and speed. Phusion DNA Polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. With an error rate > 50-fold lower than that of Taq DNA Polymerase and 6-fold lower than that of Pyrococcus furiosus DNA Polymerase (1), Phusion is one of the most accurate thermostable polymerases available. Phusion DNA Polymerase possesses 5´→ 3´ polymerase activity, 3´→ 5´ exonuclease activity and will generate blunt-ended products.

Phusion DNA Polymerase is supplied with standard 5X Phusion HF Buffer, as well as 5X Phusion GC Buffer, which can be used for complex or GC-rich templates. Each of these buffers contains MgCl2 (1.5 mM at the final [1X] reaction concentration). Reactions can also be optimized using the provided DMSO or MgCl2 solutions.

Fidelity assays were performed using a lacI-based method modified from Frey & Suppman, 1995.
A 3.8 kb fragment was amplified from 50 ng of Jurkat gDNA using different polymerases. Reactions were carried out according to the manufacturer's recommended conditions. Extension times are indicated (in minutes). Ladder L is a 1 kb DNA Ladder (NEB# N3232).
Product Source
An E. coli strain that carries the Phusion DNA Polymerase gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Phusion® High-Fidelity DNA Polymerase M0530SVIAL -20 1 x 0.05 ml 2,000 units/ml
  MgCl2 solution B0510AVIAL -20 1 x 1.5 ml 50 mM
  DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
  Phusion® HF Buffer Pack B0518SVIAL -20 2 x 1.5 ml 5 X
  Phusion® GC Buffer Pack B0519SVIAL -20 1 x 1.5 ml 5 X
  Phusion® High-Fidelity DNA Polymerase M0530LVIAL -20 1 x 0.25 ml 2,000 units/ml
  MgCl2 solution B0510AVIAL -20 2 x 1.5 ml 50 mM
  DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
  Phusion® HF Buffer Pack B0518SVIAL -20 6 x 1.5 ml 5 X
  Phusion® GC Buffer Pack B0519SVIAL -20 2 x 1.5 ml 5 X
Features
  • Robust Reactions – Maximal success with minimal optimization
  • Extreme Fidelity – >50X greater than Taq.
  • High Speed – Dramatically reduced extension times (10X faster than Pfu)
  • High Yield – Increased product yield using minimal amount of enzyme
  • Versatile – Can be used for routine PCR as well as long or difficult templates
Application Features
  • PCR
  • Cloning
  • Long or Difficult Amplification
  • High-Throughput PCR

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C.

Storage Buffer

20 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
1X stabilizers
pH 7.4 @ 25°C

Heat Inactivation

No

5' - 3' Exonuclease

No

3' - 5' Exonuclease

Yes

Unit Assay Conditions

25 mM TAPS-HCl (pH 9.3 @ 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM b-mercaptoethanol, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.

References
  • Frey, B. and Suppman, B. (1995). BioChemica. 2, 34-35.
  • Chester, N. and Marshak, D.R. (1993). Analytical Biochemistry. 209, 284-290.
Additional Citations
  • Wrenbeck E.E., Klesmith J.R., Stapleton J.A., Adeniran A., Tyo K.E., & Whitehead T.A (2016) Plasmid-based one-pot saturation mutagenesis Nat Methods 11, 928-930. DOI: 10.1038/NMETH.4029
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.Phusion® DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific. This product is manufactured by New England Biolabs, Inc. under agreement with, and under the performance specifications of Thermo Fisher Scientific.

Licenses/Patents/Disclaimers:
Notice to purchaser: Limited license.
The purchase price of this product includes a limited, non-transferable license under U.S. and foreign patents owned by BIO-RAD Laboratories, Inc., to use this product. No other license under these patents is conveyed expressly or by implication by purchase of this product.

PHUSION® is a registered trademark and property of Thermo Fisher Scientific.
THERMO SCIENTIFIC® is a registered trademark of Thermo Fisher Scientific.

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