Product Class: Kit

Phusion® High-Fidelity PCR Kit

Product Introduction

Ideal 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
  • Kit includes everything needed for a PCR reaction, including controls
Catalog # Size Concentration
E0553S 50.0 reactions
E0553L 200.0 reactions

Product Information

Description

High-Fidelity DNA Polymerases are important for applications in which the DNA sequence needs to be correct after amplification. Phusion High-Fidelity DNA Polymerase offers both high-fidelity and robust performance, and thus can be used for all PCR applifications. 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 of Taq DNA Polymerase and 6-fold lower than that of Pyrococcus furiosus DNA Polymerase (7), 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.

The Phusion High-Fidelity PCR Kit contains a sufficient supply of Phusion High-Fidelity DNA Polymerase, Phusion HF and GC Buffers, deoxynucleotides, magnesium chloride, DMSO, and DNA size standard to perform 50 reactions (small) or 200 reactions (large). Control template and primers are provided for 20 control reactions.

Notice to Customers:
Phusion® DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific. This product is now manufactured by New England Biolabs, Inc. under agreement with, and under the performance specifications of Thermo Fisher Scientific. Further details on this change can be found in the following customer communication. However, it is important to note that the catalog numbers for these products have changed. For any questions, please use our Phusion Request for Technical Support or call NEB Technical Support at 1-800-632-7799.

Phusion® is a registered trademark and property of Thermo Fisher Scientific.

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 (cycling conditions are shown below). Extension times are indicated (in minutes). Ladder L is a 1 kb DNA Ladder .
This product is related to the following categories:
Phusion® High-Fidelity DNA Polymerases Products,
This product can be used in the following applications:
Long Range PCR,
Fast PCR,
High-Fidelity PCR,
Multiplex PCR,
Specialty PCR,
Routine PCR,
PCR

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Features

Application features:
  • PCR
  • Primer Extension
  • Cloning
  • Longer Difficult Amplification
  • High-Throughput PCR

References

  1. Saiki, R.K. et al. (1985). Science. 230, 1350-1354.
  2. Chien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact.. 127, 1550-1557.
  3. Kaledin, A.S., Slyusarenko, A.G. and Gorodetskii, S.I. (1980). Biokhimiya. 45, 644-651.
  4. Lawyer, F.C. et al. (1993). PCR Method and Appl.. 2, 275-287.
  5. Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990). Nucleic Acids Res.. 18, 7317-7322.
  6. Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993). Science. 260, 778-783.
  7. Frey and Suppman (1995). Biochimica. 34-35.
  8. Breslauer, et al. (1986). PNAS. 83, 3746-3750.

Protocols, Manuals & Usage

Protocols

  1. PCR Optimization of the Control Template using Phusion® High-Fidelity PCR Kit
  2. Protocol for a Routine PCR with Phusion® High-Fidelity PCR Kit
  3. PCR Optimization with Phusion® High-Fidelity PCR Kit

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What are the differences between the various available Phusion® products?
  2. My results are not as expected. Where can I find troubleshooting help?
  3. What ends will my PCR products have?
  4. What does exonuclease activity mean for a DNA polymerase?
  5. Does Phusion® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
  6. What are the properties of this polymerase (fidelity, product ends, max amplicon, modified base incorporation, etc.)?
  7. How should I determine the appropriate annealing temperature for my reaction?
  8. What should the final primer concentration be in my PCR?
  9. My template is GC-rich or supercoiled. How can I optimize my product yield using Phusion® High-Fidelity DNA Polymerase?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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.

Licenses

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. Phusion® is a registered trademark and property of Thermo Fisher Scientific.

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 to the purchaser by the purchase of this product.

The purchase price of this product includes a limited, non-transferable license under a patent owned by New England Biolabs, Inc., to use this product. No other license under this patent is conveyed expressly or by implication to the purchaser by the purchase of this product.

QUICKLOAD® is a registered trademark of New England Biolabs, Inc.

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