Phusion® Hot Start Flex DNA Polymerase |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 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 polymerases available. Phusion DNA Polymerase possesses 5´→ 3´ polymerase activity, 3´→ 5´ exonuclease activity and will generate blunt-ended products.

Phusion Hot Start Flex DNA Polymerase offers robust, high fidelity performance and room temperature reaction setup. The addition of an aptamer-based inhibitor allows room temperature reaction setup. Phusion Hot Start Flex 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 Hot Start Flex DNA Polymerase is unlike other enzymes and care must be taken when designing cycling protocols. To determine the optimal annealing temperatures for a given set of primers, use of the NEB Tm Calculator is highly recommended.

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:
Hot Start PCR,
Long Range PCR,
Fast PCR,

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

PCR

  • 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
    • M0535S     -20    
        Phusion® Hot Start Flex DNA Polymerase M0535SVIAL -20 1 x 0.05 ml 2,000 units/ml
        Phusion® HF Buffer Pack B0518SVIAL -20 2 x 1.5 ml 5 X
        MgCl2 solution B0510AVIAL -20 1 x 1.5 ml 50 mM
        DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
        Phusion® GC Buffer Pack B0519SVIAL -20 1 x 1.5 ml 5 X
    • M0535L     -20    
        Phusion® Hot Start Flex DNA Polymerase M0535LVIAL -20 1 x 0.25 ml 2,000 units/ml
        Phusion® HF Buffer Pack B0518SVIAL -20 6 x 1.5 ml 5 X
        MgCl2 solution B0510AVIAL -20 2 x 1.5 ml 50 mM
        DMSO B0515AVIAL -20 1 x 0.5 ml 100 %
        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

    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 β-mercaptoethanol, 200 μM dNTPs including [3H]- dTTP and 15 nM primed M13 DNA. 

  • 优势和特性

    应用特性

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

  • 相关产品

    相关产品

    • dNTP 混合液
    • dNTP 套装
    • MgCl2 溶液
    • m0530-phusion-high-fidelity-dna-polymerase
    • Phusion® High-Fidelity PCR Kit
    • Phusion® 超保真 PCR 预混液（提供 HF 缓冲液）
    • Phusion® 热启动 Flex 2X 预混液
    • Phusion® 超保真 PCR 预混液（提供 GC 缓冲液）

    单独销售的组分

    • Phusion® GC 反应缓冲液套装
    • Phusion® HF 反应缓冲液套装

  • 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, 284-029.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for Phusion® Hot Start Flex DNA Polymerase (M0535)

  • 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?
    10. What are Hot Start and WarmStart® polymerases and when would I use them?

  • 问题解决指南

    • PCR Troubleshooting Guide

  • 实验技巧

    Phusion Hot Start Flex has different annealing temperature requirements than most PCR enzymes.
    Please check out the NEB Tm Calculator to help you determine your optimal Phusion annealing temperature.