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

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Phusion® High-Fidelity PCR Kit

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

Phusion® High-Fidelity PCR Kit |

库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
E0553S 不适用 50 reactions ¥679.00 有 有 有 有 有
E0553L 不适用 200 reactions ¥2,329.00 有 有 有 无 无
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. 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

    Kit 组分

    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

  • 特性和用法

  • 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

    • Deoxynucleotide (dNTP) Solution Set
    • Magnesium Chloride (MgCl2) Solution
    • Phusion® High-Fidelity PCR Master Mix with HF Buffer
    • Phusion® High-Fidelity PCR Master Mix with GC Buffer
    • Phusion® Hot Start Flex DNA Polymerase
    • Phusion® Hot Start Flex 2X Master Mix

    单独销售的组分

    • Phusion® High-Fidelity DNA Polymerase
    • Deoxynucleotide (dNTP) Solution Mix
    • 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 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. 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?
    6. My template is GC-rich or supercoiled. How can I optimize my product yield using Phusion® High-Fidelity DNA Polymerase?
    7. What are the advantages to using Phusion® High-Fidelity DNA Polymerase?
    8. What is the error rate of Phusion® High-Fidelity DNA Polymerase?
    9. 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?
    10. Why do I see no product or low yield on an agarose gel after a PCR using Phusion® High-Fidelity DNA Polymerase?
    11. 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?
    12. Will Phusion® High-Fidelity DNA Polymerase incorporate dUTPs?
    13. Why are there low molecular weight discrete bands on an agarose gel after a PCR using Phusion® High-Fidelity DNA Polymerase?
    14. I’d like to clone a fragment amplified with Phusion® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
    15. What should my primer concentration be when using Phusion® High-Fidelity DNA Polymerase/Phusion® High-Fidelity PCR Master Mix?

  • 问题解决指南
    • PCR Troubleshooting Guide

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Phusion® High-Fidelity PCR Kit | 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

    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]

    • E0553S_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#]

    • E0553S_L_v1_0151708
    • E0553S_v1_10013495
    • E0553L_v1_10013926
    • E0553S_v1_10021738
    • E0553L_v1_10021732
    • E0553S_v1_10023549
    • E0553L_v1_10034492
    • E0553S_v1_10034491
    • E0553L_v1_10041340
    • E0553S_v1_10041432
    • E0553L_v1_10045328
    • E0553S_v1_10047984
    • E0553L_v1_10048545
    • E0553L_v1_10050935
    • E0553S_v1_10054175
    • E0553S_v1_10055991
    • E0553L_v1_10055985
    • E0553L_v1_10060090
    • E0553S_v1_10061490
    • E0553L_v1_10063721
    • E0553S_v1_10064331
    • E0553S_v1_10065626
    • E0553L_v1_10069926
    • E0553S_v1_10073566
    • E0553L_v1_10077741
    • E0553S_v1_10079225
    • E0553L_v1_10082375
    • E0553L_v1_10084849
    • E0553L_v1_10086366
    • E0553S_v1_10086370
    • E0553L_v1_10090571
    • E0553S_v1_10091455
    • E0553L_v1_10095925
    • E0553S_v1_10095929
    • E0553L_v1_10099933
    • E0553S_v1_10099932
    • E0553L_v1_10104358
    • E0553S_v1_10104881
    • E0553L_v1_10107125
    • E0553S_v1_10108362
    • E0553L_v1_10109593
    • E0553S_v1_10113742
    • E0553L_v1_10117360
    • E0553S_v1_10115049
    • E0553S_v1_10120996
    • E0553L_v1_10120995
    • E0553L_v1_10123923
    • E0553L_v1_10125446
    • E0553L_v1_10131931
    • E0553L_v1_10132254
    • E0553S_v1_10127745
    • E0553S_v1_10137416
    • E0553L_v1_10140019
    • E0553L_v1_10145819
    • E0553S_v1_10145487
    • E0553L_v1_10150541
    • E0553S_v1_10150731
    • E0553L_v1_10153111
    • E0553S_v1_10157016
    • E0553L_v1_10157024
    • E0553S_v1_10161330
    • E0553L_v1_10161329

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Phusion® High-Fidelity DNA Polymerase
    • Phusion® HF Buffer Pack
    • Phusion® GC Buffer Pack
    • Deoxynucleotide (dNTP) Solution Mix
    • MgCl2 solution
    • Control Lambda Template
    • 1.3 kb Control Primer Mix
    • 10 kb Control Primer Mix
    • DMSO
    • Quick-Load DNA Marker, Broad Range