Phusion® Hot Start Flex DNA Polymerase | NEB酶试剂 New England Biolabs

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Phusion® Hot Start Flex 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 with minimal optimization
  •  Hot start formulation enables room temperature reaction setup and reduces non-specific amplification
  • High yields with shorter extension times

Phusion® Hot Start Flex DNA Polymerase | NEB酶试剂 New England Biolabs

库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0535S 2,000 units/ml 100 units ¥1,309.00 有 有 有 无 有
M0535L 2,000 units/ml 500 units ¥5,219.00 有 有 无 无 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. 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

    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

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

    单独销售的组分

    • Phusion® GC Buffer Pack
    • Phusion® HF 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, 284-029.

操作说明、说明书 & 用法

  • 操作说明
    1. PCR 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. Is Phusion® Hot Start Flex DNA Polymerase the same as Phusion Hot Start DNA Polymerase (#F-540S/L) or Phusion Hot Start II DNA Polymerase (#F-549S/L), which were previously sold by NEB?
    2. Are the DNA fragments products by Phusion® Hot Start Flex DNA Polymerase blunt-ended or do they have the single-base 3’ overhang that Taq DNA Polymerase yields?
    3. What are the advantages of using Phusion® Hot Start Flex DNA Polymerase?
    4. I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Phusion® Hot Start Flex DNA Polymerase?
    5. Why do I see no product or low yield on an agarose gel after a PCR using Phusion® Hot Start Flex DNA Polymerase?
    6. Will Phusion® Hot Start Flex DNA Polymerase incorporate dUTPs?
    7. Why are there low molecular weight discrete bands on an agarose gel after a PCR using Phusion® Hot Start Flex DNA Polymerase?
    8. I’d like to clone a fragment amplified with Phusion® Hot Start Flex DNA Polymerase. Do I have to blunt-end clone?
    9. I see foaming when my Phusion® Hot Start Flex-amplified PCR products are spotted on microarray slides.
    10. What should my primer concentration be when using Phusion® Hot Start Flex DNA Polymerase?
    11. How does the fidelity of Phusion® DNA Polymerase compare to Taq DNA Polymerase?
    12. Why are there high molecular weight smears or DNA in the wells of an agarose gel after a PCR using Phusion Hot Start Flex DNA Polymerase?
    13. Does Phusion® Hot Start Flex DNA Polymerase exhibit a strand displacement activity?
    14. How do I activate Phusion® Hot Start Flex DNA Polymerase?

  • 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.

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Phusion® Hot Start Flex DNA Polymerase | NEB酶试剂 New England Biolabs 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?
    • Using aptamers to control enzyme activities: Hot Start Taq and beyond

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]

    • M0535S_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#]

    • M0535S_L_v1_0041509
    • M0535S_L_v1_0041602
    • M0535S_L_v1_0041608
    • M0535S_L_v1_0061704
    • M0535S_L_v1_0071707
    • M0535S_L_v1_0071709
    • M0535S_L_v1_0071803
    • M0535L_v1_10010941
    • M0535S_v1_10015640
    • M0535L_v1_10015642
    • M0535L_v1_10020353
    • M0535S_v1_10020361
    • M0535L_v1_10022721
    • M0535S_v1_10029410
    • M0535L_v1_10030772
    • M0535S_v1_10034493
    • M0535S_v1_10038930
    • M0535L_v0_10038929
    • M0535L_v1_10046508
    • M0535S_v1_10046509
    • M0535L_v1_10047942
    • M0535S_v1_10047941
    • M0535L_v1_10050376
    • M0535S_v1_10053292
    • M0535L_v1_10054145
    • M0535S_v1_10056369
    • M0535L_v1_10057283
    • M0535L_v1_10063082
    • M0535S_v1_10063684
    • M0535L_v1_10069245
    • M0535L_v1_10071789
    • M0535S_v1_10071792
    • M0535L_v1_10072586
    • M0535S_v1_10077371
    • M0535S_v1_10079475
    • M0535L_v1_10075653
    • M0535S_v1_10082802
    • M0535L_v1_10085127
    • M0535S_v1_10088393
    • M0535L_v1_10087289
    • M0535S_v1_10097880
    • M0535L_v1_10094242
    • M0535S_v1_10101815
    • M0535L_v1_10104082
    • M0535S_v1_10108148
    • M0535L_v1_10110277
    • M0535S_v1_10109311
    • M0535L_v1_10115657
    • M0535S_v1_10115038
    • M0535L_v1_10118877
    • M0535S_v1_10119087
    • M0535L_v1_10127255
    • M0535L_v1_10129795
    • M0535S_v1_10129537
    • M0535L_v1_10135393
    • M0535L_v1_10140649
    • M0535S_v1_10139177
    • M0535L_v1_10146959
    • M0535S_v1_10147110
    • M0535L_v1_10151792
    • M0535S_v1_10151857
    • M0535S_v1_10157017
    • M0535L_v1_10158253

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Phusion® Hot Start Flex DNA Polymerase
    • Phusion® HF Buffer Pack
    • MgCl2 solution
    • DMSO
    • Phusion® GC Buffer Pack