Phusion® High-Fidelity PCR Kit |NEB酶试剂 New England Biolabs

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产品信息

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.

Phusion® High-Fidelity PCR Kit |
Fidelity assays were performed using a lacI-based method modified from Frey & Suppman, 1995.

Phusion® High-Fidelity PCR Kit |
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 .
产品类别:
Phusion® High-Fidelity DNA Polymerases Products

应用:
Long Range PCR,
Fast PCR,
High-Fidelity PCR,

Multiplex PCR,
Specialty PCR,
Routine PCR,

PCR

  • 试剂盒组成

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • E0553S     -20    
        Phusion® High-Fidelity DNA Polymerase M0530AVIAL -20 1 x 0.025 ml 2,000 units/ml
        Phusion® HF Buffer Pack B0518SVIAL -20 1 x 1.5 ml 5 X
        Phusion® GC Buffer Pack B0519SVIAL -20 1 x 1.5 ml 5 X
        Deoxynucleotide (dNTP) Solution Mix N0447AVIAL -20 1 x 0.1 ml 10 mM each dNTP
        MgCl2 solution B0510AVIAL -20 1 x 1.5 ml 50 mM
        Control Lambda Template N3010AVIAL -20 1 x 0.04 ml 0.5 ng/µl
        1.3 kb Control Primer Mix S0535AVIAL -20 1 x 0.05 ml 4 µM
        10 kb Control Primer Mix S0536AVIAL -20 1 x 0.05 ml 4 µM
        DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
        Quick-Load DNA Marker, Broad Range N0303AVIAL -20 1 x 0.2 ml 100 µg/ml
    • E0553L     -20    
        Phusion® High-Fidelity DNA Polymerase M0530AAVIAL -20 1 x 0.1 ml 2,000 units/ml
        Phusion® HF Buffer Pack B0518SVIAL -20 3 x 1.5 ml 5 X
        Phusion® GC Buffer Pack B0519SVIAL -20 1 x 1.5 ml 5 X
        Deoxynucleotide (dNTP) Solution Mix N0447AAVIAL -20 1 x 0.4 ml 10 mM each dNTP
        MgCl2 solution B0510AVIAL -20 1 x 1.5 ml 50 mM
        Control Lambda Template N3010AVIAL -20 1 x 0.04 ml 0.5 ng/µl
        1.3 kb Control Primer Mix S0535AVIAL -20 1 x 0.05 ml 4 µM
        10 kb Control Primer Mix S0536AVIAL -20 1 x 0.05 ml 4 µM
        DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
        Quick-Load DNA Marker, Broad Range N0303AAVIAL -20 1 x 0.4 ml 100 µg/ml

  • 优势和特性

    Features

    Application features:

    • PCR
    • Primer Extension
    • Cloning
    • Longer Difficult Amplification
    • High-Throughput PCR

  • 相关产品

    相关产品

    • dNTP 套装
    • MgCl2 溶液
    • Phusion® 超保真 PCR 预混液（提供 HF 缓冲液）
    • Phusion® 超保真 PCR 预混液（提供 GC 缓冲液）
    • Phusion® Hot Start Flex DNA Polymerase
    • Phusion® 热启动 Flex 2X 预混液

    单独销售的组分

    • m0530-phusion-high-fidelity-dna-polymerase
    • dNTP 混合液
    • Lambda DNA

  • 参考文献

    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.

操作说明、说明书 & 用法

  • 操作说明

    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

  • 说明书

    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE0553

  • 使用指南

    • Activity of Restriction Enzymes in PCR Buffers
    • Guidelines for PCR Optimization with Thermophilic DNA Polymerases

工具 & 资源

  • 选择指南

    • DNA Polymerase Selection Chart
    • Thermophilic DNA Polymerases

  • Web 工具

    • Tm Calculator

FAQs & 问题解决指南

  • 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?

  • 问题解决指南

    • PCR Troubleshooting Guide