Phusion® GC 反应缓冲液套装 |NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品信息

NEB 公司提供专用的 5X 反应缓冲液和补充试剂,适用于 Thermo Scientific® Phusion 超保真 DNA 聚合酶及相关产品;缓冲液和试剂均由 NEB 公司生产并进行质量控制。

产品类别:
Phusion® High-Fidelity DNA Polymerases Products,
Reaction Buffers Products,
Buffers 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
    • B0519S     -20    
        Phusion® GC Buffer Pack B0519SVIAL -20 4 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 %

  • 相关产品

    相关产品

    • Phusion® HF 反应缓冲液套装
    • Phusion® High-Fidelity PCR Kit
    • m0530-phusion-high-fidelity-dna-polymerase
    • Phusion® 超保真 PCR 预混液(提供 HF 缓冲液)
    • Phusion® 超保真 PCR 预混液(提供 GC 缓冲液)
    • Phusion® Hot Start Flex DNA Polymerase
    • Phusion® 热启动 Flex 2X 预混液

操作说明、说明书 & 用法

  • 使用指南

    • Activity of Restriction Enzymes in PCR Buffers

工具 & 资源

  • Web 工具

    • Tm Calculator

FAQs & 问题解决指南

  • FAQs

    1. What are the differences between the various available Phusion® products?

Phusion® HF 反应缓冲液套装 |NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品信息

NEB 公司提供专用的 5X 反应缓冲液和补充试剂,适用于 Thermo Scientific® Phusion 超保真 DNA 聚合酶及相关产品;缓冲液和试剂均由 NEB 公司生产并进行质量控制。

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

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

High-Fidelity 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
    • B0518S     -20    
        Phusion® HF Buffer Pack B0518SVIAL -20 4 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 %

  • 相关产品

    相关产品

    • Phusion® GC 反应缓冲液套装
    • Phusion® High-Fidelity PCR Kit
    • m0530-phusion-high-fidelity-dna-polymerase
    • Phusion® 超保真 PCR 预混液(提供 HF 缓冲液)
    • Phusion® 超保真 PCR 预混液(提供 GC 缓冲液)
    • Phusion® Hot Start Flex DNA Polymerase
    • Phusion® 热启动 Flex 2X 预混液

操作说明、说明书 & 用法

  • 使用指南

    • Activity of Restriction Enzymes in PCR Buffers

工具 & 资源

  • Web 工具

    • Tm Calculator

FAQs & 问题解决指南

  • FAQs

    1. What are the differences between the various available Phusion® products?

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

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

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

    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

    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

    • dNTP 套装
    • dNTP 混合液
    • MgCl2 溶液
    • Phusion® High-Fidelity PCR Kit
    • Phusion® 超保真 PCR 预混液(提供 HF 缓冲液)
    • Phusion® 超保真 PCR 预混液(提供 GC 缓冲液)
    • dNTP 混合液
    • Phusion® HF 反应缓冲液套装
    • Phusion® GC 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, 284-290.

操作说明、说明书 & 用法

  • 操作说明

    1. PCR Protocol for Phusion® High-Fidelity DNA Polymerase (M0530)

  • 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?

  • 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.

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

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

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

    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.