Phusion® High-Fidelity DNA Polymerase |NEB酶试剂 New England Biolabs

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Phusion® High-Fidelity DNA Polymerase

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

Phusion® High-Fidelity DNA Polymerase |

库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0530S 2,000 units/ml 100 units ¥1,029.00 有 有 有 有 有
M0530L 2,000 units/ml 500 units ¥4,109.00 有 有 有 有 有
如何订购
Please enter a quantity for at least one size

产品信息

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.

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

产品来源

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

产品类别:
Phusion® High-Fidelity DNA Polymerases Products

应用:
Gibson Assembly®,
Long Range PCR,
Fast PCR,

High-Fidelity PCR,
Multiplex PCR,
Specialty PCR,
Routine PCR,

PCR

  • 产品组分信息

    产品组分信息

    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
    • M0530S     -20    
        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
    • M0530L     -20    
        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

  • 特性和用法

    单位定义

    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.

    贮存溶液

    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

    热失活

    否

    5′ – 3′ 核酸外切酶

    No

    3′ – 5′ 核酸外切酶

    Yes

    单位活性检测条件

    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.

  • 优势和特性

    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

    应用特性

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

  • 相关产品

    相关产品

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

  • 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. PCR Protocol Phusion® DNA Polymerase

  • 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. Which thermophilic DNA polymerase should I use?
    4. Can I continue to use the same reaction conditions if I use Phusion® products with the new product numbers?
    5. Are the prices and sizes of these products the same now that they are manufactured by NEB?
    6. 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?
    7. My template is GC-rich or supercoiled. How can I optimize my product yield using Phusion® High-Fidelity DNA Polymerase?
    8. What are the advantages of using Phusion® Hot Start Flex DNA Polymerase?
    9. What is the error rate of Phusion® High-Fidelity DNA Polymerase?
    10. 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?
    11. Why do I see no product or low yield on an agarose gel after a PCR using Phusion® High-Fidelity DNA Polymerase?
    12. 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?
    13. Will Phusion® High-Fidelity DNA Polymerase incorporate dUTPs?
    14. Why are there low molecular weight discrete bands on an agarose gel after a PCR using Phusion® High-Fidelity DNA Polymerase?
    15. I’d like to clone a fragment amplified with Phusion® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
    16. I see foaming when my Phusion® High-Fidelity DNA Polymerase-amplified PCR products are spotted on microarray slides.
    17. What should my primer concentration be when using Phusion® High-Fidelity DNA Polymerase/Phusion® High-Fidelity PCR Master Mix?
    18. I am competing in the iGEM competition. Do you have any products that I should consider from NEB?

  • 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 NEB’s Tm Calculator to help you determine your optimal Phusion annealing temperature.

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Phusion® High-Fidelity DNA Polymerase | Powered by Bioz See more details on Bioz

    更多引用文献

    • 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

  • 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

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]

    • M0530S_L_v1

  • 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#]

    • M0530S_L_v1_0051509
    • M0530S_L_v1_0051503
    • M0530S_L_v1_0051603
    • M0530S_L_v1_0051604
    • M0530S_L_v1_0051609
    • M0530S_L_v1_0051704
    • M0530S_L_v1_0051708
    • M0530S_L_v1_0051803
    • M0530L_v1_10010610
    • M0530S_v1_10010607
    • M0530L_v1_10012019
    • M0530S_v1_10019998
    • M0530L_v1_10023482
    • M0530S_v1_10023480
    • M0530L_v1_10027462
    • M0530S_v1_10027455
    • M0530L_v1_10033270
    • M0530S_v1_10033269
    • M0530L_v1_10038569
    • M0530S_v1_10038568
    • M0530L_v1_10045618
    • M0530S_v1_10045600
    • M0530L_v1_10054618
    • M0530S_v1_10054614
    • M0530L_v1_10062230
    • M0530S_v1_10062227
    • M0530L_v1_10073256
    • M0530L_v1_10076890
    • M0530S_v1_10079674
    • M0530L_v1_10079675
    • M0530L_v1_10083326
    • M0530S_v1_10083325
    • M0530L_v1_10087074
    • M0530S_v1_10087076
    • M0530L_v1_10090934
    • M0530S_v1_10090926
    • M0530L_v1_10095930
    • M0530S_v1_10095932
    • M0530S_v1_10099958
    • M0530L_v1_10099959
    • M0530S_v1_10104095
    • M0530L_v1_10104208
    • M0530S_v1_10107234
    • M0530L_v1_10107238
    • M0530S_v1_10108878
    • M0530L_v1_10111589
    • M0530S_v1_10114107
    • M0530L_v1_10111588
    • M0530S_v1_10115894
    • M0530L_v1_10119089
    • M0530S_v1_10121887
    • M0530L_v1_10122326
    • M0530L_v1_10125448
    • M0530L_v1_10129951
    • M0530S_v1_10125467
    • M0530L_v1_10132260
    • M0530S_v1_10132266
    • M0530L_v1_10137982
    • M0530S_v1_10137986
    • M0530S_v1_10139549
    • M0530L_v1_10140979
    • M0530S_v1_10143131
    • M0530S_v1_10145976
    • M0530L_v1_10147412
    • M0530S_v1_10147197
    • M0530L_v1_10148882
    • M0530S_v1_10151105
    • M0530L_v1_10151560
    • M0530L_v1_10155193
    • M0530S_v1_10153547
    • M0530L_v1_10157026
    • M0530S_v1_10160670

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Phusion® High-Fidelity DNA Polymerase
    • MgCl2 solution
    • DMSO
    • Phusion® HF Buffer Pack
    • Phusion® GC Buffer Pack