Phusion® High-Fidelity PCR Master Mix with HF Buffer |NEB酶试剂 New England Biolabs

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Phusion® High-Fidelity PCR Master Mix with HF Buffer

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 compared to Taq
  • Robust reactions – maximal success with minimal optimization
  • Master mix format with HF buffer ideal for a wide range of AT/GC content

Phusion® High-Fidelity PCR Master Mix with HF Buffer |

货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0531S 2 X 100 reactions (50 µl) ¥1,759.00
M0531L 2 X 500 reactions (50 µl) ¥6,789.00
Please enter a quantity for at least one size


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 applications. 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 polymerase available. Phusion DNA Polymerase possesses 5´→ 3´ polymerase activity, 3´→ 5´ exonuclease activity and will generate blunt-ended products.

Phusion High-Fidelity PCR Master Mix with HF Buffer is a 2X master mix consisting of Phusion DNA Polymerase, deoxynucleotides and reaction buffer that has been optimized and includes MgCl2. All that is required is the addition of template, primers and water.

Phusion® High-Fidelity PCR Master Mix with HF Buffer |
Fidelity assays were performed using a lacI-based method modified from Frey & Suppman, 1995.
Phusion® High-Fidelity PCR Master Mix with HF Buffer |
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.


An E. coli strain that carries the Phusion DNA Polymerase gene.

Phusion® High-Fidelity DNA Polymerases Products,
Master Mixes Products

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

Multiplex PCR,
Specialty PCR,
Routine PCR,


  • 产品组分信息



    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0531S     -20    
        Phusion® High-Fidelity PCR Master Mix with HF Buffer M0531SVIAL -20 2 x 1.25 ml 2 X
        DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
    • M0531L     -20    
        Phusion® High-Fidelity PCR Master Mix with HF Buffer M0531SVIAL -20 10 x 1.25 ml 2 X
        DMSO B0515AVIAL -20 2 x 0.5 ml 100 %

  • 特性和用法



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

  • 优势和特性


    • 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 per as well as long or difficult templates.


    • PCR
    • Cloning
    • Long or Difficult Amplification
    • High-Throughput PCR

  • 相关产品


    • Deoxynucleotide (dNTP) Solution Mix
    • Deoxynucleotide (dNTP) Solution Set
    • Magnesium Chloride (MgCl2) Solution
    • Phusion® GC Buffer Pack
    • Phusion® HF Buffer Pack
    • Phusion® High-Fidelity DNA Polymerase
    • Phusion® High-Fidelity PCR Kit
    • Phusion® High-Fidelity PCR Master Mix with GC Buffer

  • 注意事项
    1. Products specifications for individual components in the Phusion High-Fidelity PCR Master Mix with HF Buffer are available separately.

  • 参考文献
    1. Frey, B. and Suppman, B. (1995). Biochemica. 2, 34-35.
    2. Chester, N. and Marshak, D.R. (1993). Analytical Biochemistry. 209, 284-290.

操作说明、说明书 & 用法

  • 操作说明
    1. Protocol Phusion® High-Fidelity PCR Master Mix with HF Buffer

  • 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 has changed?
    2. Have the formulations or any other characteristics of these products changed now that they are manufactured by NEB?
    3. Can I continue to use the same reaction conditions if I use Phusion® products with the new product numbers?
    4. Are the prices and sizes of these products the same now that they are manufactured by NEB?
    5. What are the advantages to using Phusion® High-Fidelity PCR Master Mixes?
    6. Why are there low molecular weight discrete bands on an agarose gel after a PCR using Phusion® High-Fidelity PCR Master Mix?
    7. Does Phusion® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
    8. What should my primer concentration be when using Phusion® High-Fidelity DNA Polymerase/Phusion® High-Fidelity PCR Master Mix?
    9. Will Phusion® High-Fidelity DNA Polymerase incorporate dUTPs?
    10. Why are there high molecular weight smears or DNA in the wells of an agarose gel after a PCR using Phusion® High-Fidelity DNA Polymerase?
    11. Are the DNA fragments produced by Phusion® High-Fidelity DNA Polymerase blunt-ended or do they have the single-base 3´ overhang that Taq DNA Polymerase yields?
    12. What is the error rate of Phusion® High-Fidelity DNA Polymerase?
    13. I’d like to clone a fragment amplified with Phusion® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
    14. I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Phusion® High-Fidelity DNA Polymerase?

  • 问题解决指南
    • PCR Troubleshooting Guide

  • 实验技巧
    Phusion has different annealing temperature requirements than most PCR enzymes. Please check out to help you determine your optimal Phusion annealing temperature.

引用 & 技术文献

  • 引用文献


    Phusion® High-Fidelity PCR Master Mix with HF Buffer | Powered by Bioz See more details on Bioz

  • 专题文章
    • Polymerase Fidelity: What is it, and what does it mean for your PCR?

质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控,以满足指定的质量标准,对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息,可从此处查阅。

  • 产品说明与变更通知



    • M0531S_L_v2

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控,这些文件的命名规则如下: [货号]_[规格]_[版本]_[Lot#]

    • M0531S_L_v2_0281610
    • M0531S_L_v2_0301706
    • M0531S_L_v2_0291704
    • M0531S_L_v2_0311709
    • M0531S_L_v2_0321712
    • M0531L_v2_10014771
    • M0531S_v2_10013407
    • M0531L_v1_10017665
    • M0531S_v1_10017666
    • M0531L_v2_10021846
    • M0531L_v2_10024141
    • M0531L_v2_10029414
    • M0531S_v2_10024142
    • M0531S_v2_10031782
    • M0531L_v2_10034149
    • M0531S_v2_10034173
    • M0531L_v2_10037070
    • M0531S_v2_10038923
    • M0531S_v2_10043495
    • M0531L_v2_10044067
    • M0531L_v2_10047035
    • M0531S_v2_10047034
    • M0531L_v2_10049316
    • M0531S_v2_10049318
    • M0531S_v2_10050375
    • M0531L_v2_10051774
    • M0531S_v2_10053732
    • M0531L_v2_10056074
    • M0531S_v2_10056076
    • M0531L_v2_10057799
    • M0531S_v2_10061441
    • M0531S_v2_10065627
    • M0531L_v2_10064096
    • M0531S_v2_10069242
    • M0531L_v2_10073033
    • M0531S_v2_10077104
    • M0531L_v2_10080030
    • M0531L_v2_10082447
    • M0531S_v2_10085028
    • M0531L_v2_10085029
    • M0531S_v2_10088144
    • M0531L_v2_10090727
    • M0531S_v2_10092958
    • M0531L_v2_10094789
    • M0531S_v2_10096452
    • M0531L_v2_10098898
    • M0531S_v2_10100862
    • M0531L_v2_10102885
    • M0531S_v2_10102886
    • M0531L_v2_10109652
    • M0531S_v2_10110296
    • M0531S_v2_10116313
    • M0531L_v2_10117359
    • M0531S_v2_10119823
    • M0531L_v2_10122329
    • M0531L_v2_10126807
    • M0531S_v2_10126057
    • M0531L_v2_10129529
    • M0531S_v2_10135765
    • M0531S_v2_10137919
    • M0531L_v2_10135764
    • M0531L_v2_10139171
    • M0531S_v2_10141029
    • M0531L_v2_10145389
    • M0531S_v2_10147610
    • M0531L_v2_10149745
    • M0531S_v2_10151641
    • M0531L_v2_10157958
    • M0531S_v2_10159517
    • M0531L_v2_10157005
    • M0531L_v2_10163430

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Phusion® High-Fidelity PCR Master Mix with HF Buffer
    • DMSO