Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs

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Q5® Hot Start High-Fidelity 2X Master Mix

Q5® High-Fidelity DNA Polymerase sets a new standard for both fidelity and robust performance. With the highest fidelity amplification available (~280 times higher than Taq), Q5 DNA Polymerase results in ultra-low error rates. Q5 DNA Polymerase is composed of a novel polymerase that is fused to the processivity-enhancing Sso7d DNA binding domain, improving speed, fidelity and reliability of performance.

Working with uracil-containing DNA templates or using dUTP? Learn about Q5U Hot Start High-Fidelity DNA Polymerase (NEB #M0515).

  • Highest fidelity amplification (~280X higher than Taq)
  • Ultra-low error rates
  • Superior performance for a broad range of amplicons (from high AT to high GC)
  • Bulk format available: M0494X = 1 x 12.5 ml bottle

1. Josh Quick 2020. nCoV-2019 sequencing protocol v3 (LoCost). protocols.io https://www.protocols.io/view/ncov-2019-sequencing-protocol-v3-locost-bh42j8ye


Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs

Important Tips for Q5® Polymerase


货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0494S 2 X 100 reactions ¥1,819.00
M0494L 2 X 500 reactions ¥7,249.00
M0494X 2 X 500 reactions ¥7,249.00
Please enter a quantity for at least one size

Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs Want to try this product? Request a sample Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs Need a custom/large volume order? Contact Us

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Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs

The Q5® Hot Start High-Fidelity 2X Master Mix features a high-fidelity, thermostable, hot start DNA polymerase with 3´→ 5´ exonuclease activity, fused to a processivity-enhancing Sso7d domain to support robust DNA amplification. The addition of an aptamer-based inhibitor allows room temperature reaction setup. With an error rate ~280-fold lower than that of Taq DNA Polymerase, Q5 Hot Start High-Fidelity 2X Master Mix is ideal for cloning and can be used for long or difficult amplicons. The convenient master mix formulation is supplied at a 2X concentration. The mix contains dNTPs, Mg++ and a proprietary broad-use buffer requiring only the addition of primers and DNA template for robust amplification regardless of GC content. When used at the recommended 1X final concentration, the Q5 Hot Start High-Fidelity Master Mix contains 2 mM Mg++. Q5 Hot Start High-Fidelity 2X Master Mix is unlike typical, lower fidelity PCR enzymes. To determine the optimal annealing temperatures for a given set of primers, use of the NEB Tm Calculator is highly recommended.

Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs
Q5-High Fidelity 2X Master Mix formats allow robust amplification of a broad range of targets with a single formulation. 

Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs
Amplification of a variety of human genomic amplicons from low to high GC content using Q5 Hot Start High-Fidelity 2X Master Mix. All reactions were set up at room temperature, conducted using 30 cycles of amplification and visualized by microfluidic LabChip® analysis.

Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs


An E. coli strain that carries the Q5 High-Fidelity DNA Polymerase gene.

Q5® High-Fidelity DNA Polymerases Products,
Master Mixes Products

Site Directed Mutagenesis,
Site Directed Mutagenesis,
Hot Start PCR,

Long Range PCR,
Fast PCR,
High-Fidelity PCR,
Multiplex PCR,
DNA Amplification, PCR & qPCR,
Specialty PCR,
Routine PCR,

Fast Cloning: Accelerate your cloning workflows with reagents from NEB

  • 产品组分信息



    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0494S     -20    
        Q5® Hot Start High-Fidelity 2X Master Mix M0494SVIAL -20 2 x 1.25 ml 2 X
    • M0494L     -20    
        Q5® Hot Start High-Fidelity 2X Master Mix M0494SVIAL -20 10 x 1.25 ml 2 X
    • M0494X     -20    
        Q5® Hot Start High-Fidelity 2X Master Mix M0494XVIAL -20 1 x 12.5 ml 2 X

  • 特性和用法


  • 优势和特性


    • High-specificity PCR
    • High-fidelity PCR
    • Cloning
    • Long or Difficult Amplification
    • High-throughput PCR

  • 相关产品


    • Deoxynucleotide (dNTP) Solution Mix
    • Deoxynucleotide (dNTP) Solution Set
    • Q5® High-Fidelity DNA Polymerase
    • Q5® Hot Start High-Fidelity DNA Polymerase
    • Q5® High-Fidelity 2X Master Mix
    • Q5® Reaction Buffer Pack

  • 注意事项
    1. A precipitate (most noticeable after the first 1–2 freeze/thaw cycles) is not uncommon. To ensure optimal performance, the master mix should be thawed and resuspended prior to use. Stability testing using up to 15 freeze/thaw cycles has shown no negative effect on master mix performance.
    2. Product specifications for individual components in Q5 Hot Start High-Fidelity 2X Master Mix are available separately.

  • 参考文献
    1. Anastassia Voronova Erin Coyne, Ashraf Al Madhoun, Joel V. Fair, Neven Bosiljcic, Catherine St-Louis, Grace Li, Sherry Thurig, Valerie A. Wallace, Nadine Wiper-Bergeron, and Ilona S. Skerjanc. (2013). Hedgehog Signaling Regulates MyoD Expression and Activity. J Biol Chem. 288(6), 4389–4404. PubMedID: PMC3567689
    2. Lieve Naesens, Luke Guddat, Dianne Keough, André B.P. van Kuilenburg, Judith Meijer, Johan Vande Voorde and Jan Balzarini (2013). ROLE OF HUMAN HYPOXANTHINE GUANINE PHOSPHORIBOSYLTRANSFERASE IN ACTIVATION OF THE ANTIVIRAL AGENT T-705 (FAVIPIRAVIR). Molecular Pharmacology Fast Forward. 87247,
    3. Hicham Bouabe, Klaus Okkenhaug A Protocol for Construction of Gene Targeting Vectors and Generation of Homologous Recombinant Embryonic Stem Cells. Methods in Molecular Biology .

操作说明、说明书 & 用法

  • 操作说明
    1. Protocol for Q5® Hot Start High-Fidelity 2X Master Mix

  • 使用指南
    • Activity of Restriction Enzymes in PCR Buffers
    • Guidelines for PCR Optimization with Thermophilic DNA Polymerases

  • 应用实例
    • AppNote_Multiplex_PCR_Using_Q5_HF_DNA_Polymerase

工具 & 资源

  • 选择指南
    • DNA Polymerase Selection Chart

  • Web 工具
    • Tm Calculator

FAQs & 问题解决指南

  • FAQs
    1. What are the advantages to using Q5® Hot Start High-Fidelity 2X Master Mix?
    2. What is the fidelity of Q5® High-Fidelity DNA Polymerase?
    3. How should I determine an appropriate annealing temperature for my reaction?
    4. What should my primer concentration be when using Q5® High-Fidelity DNA Polymerase products?
    5. How should I set up a PCR reaction using Q5® Hot Start 2X Master Mix?
    6. My template is GC rich or supercoiled. How can I optimize my product yield using Q5® High-Fidelity Master Mixes?
    7. There is a precipitate in the bottom of the Master Mix tube? Is this normal?
    8. How do I activate Q5® Hot Start High-Fidelity DNA Polymerase?
    9. Are the DNA fragments produced by Q5® High-Fidelity DNA Polymerase blunt-ended or do they have the single-base 3´ overhang that Taq DNA Polymerase yields?
    10. What length of product can be made by Q5® High-Fidelity DNA Polymerase?
    11. I have a tube of the Q5® High GC Enhancer from another product formulation – can I add it to the Q5 Master Mix?
    12. What are the stability and storage requirements of the Q5® Master Mixes?
    13. Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
    14. Where can I find help troubleshooting my PCR?
    15. I’d like to clone a fragment amplified with Q5® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
    16. Will Q5® High-Fidelity DNA Polymerase incorporate dUTPs?

  • 问题解决指南
    • PCR Troubleshooting Guide

引用 & 技术文献

  • 引用文献


    Q5® Hot Start High-Fidelity 2X Master Mix | NEB酶试剂 New England Biolabs Powered by Bioz See more details on Bioz

  • 出版物
    • Thuronyi, B.W., Koblan, L.W., Levy, J.M. et al (2019) Continuous evolution of base editors with expanded target compatibility and improved activity Nat Biotechnol; 37, 1070-1079.. DOI: 10.1038/s41587-019-0193-0

  • 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



    • M0494S_L_v1
    • M0494S_L_X_v2

  • 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#]

    • M0494S_L_v1_0181605
    • M0494S_L_v1_0241705
    • M0494S_L_v1_0221702
    • M0494S_L_v1_0251709
    • M0494S_L_v1_0261709
    • M0494S_L_v1_0271712
    • M0494S_L_v1_0281804
    • M0494L_v1_10010240
    • M0494L_v1_10014231
    • M0494S_v1_10013384
    • M0494L_v1_10017445
    • M0494S_v0_10016241
    • M0494S_v1_10016241
    • M0494S_v1_10022723
    • M0494X_v0_10022907
    • M0494L_v1_10022724
    • M0494L_v1_10026520
    • M0494S_v1_10026868
    • M0494S_v1_10029317
    • M0494L_v1_10029427
    • M0494S_v1_10032699
    • M0494L_v1_10032779
    • M0494S_v1_10034038
    • M0494L_v1_10035174
    • M0494S_v1_10034626
    • M0494L_v1_10036875
    • M0494S_v1_10038469
    • M0494L_v1_10041877
    • M0494S_v1_10044987
    • M0494X_v1_10038492
    • M0494L_v1_10044988
    • M0494X_v1_10047139
    • M0494L_v1_10047978
    • M0494S_v1_10046047
    • M0494X_v1_10047979
    • M0494L_v1_10049892
    • M0494L_v1_10051680
    • M0494S_v1_10050343
    • M0494S_v1_10052183
    • M0494L_v1_10052189
    • M0494X_v1_10053899
    • M0494X_v1_10054946
    • M0494S_v1_10055348
    • M0494L_v1_10055349
    • M0494L_v1_10058116
    • M0494S_v1_10057545
    • M0494X_v1_10058346
    • M0494X_v1_10060006
    • M0494L_v1_10061420
    • M0494S_v1_10060117
    • M0494L_v1_10063458
    • M0494G_v0_10064701
    • M0494S_v1_10063547
    • M0494X_v1_10022907
    • M0494X_v1_10063539
    • M0494L_v1_10066872
    • M0494S_v1_10068707
    • M0494X_v1_10068376
    • M0494L_v1_10072582
    • M0494X_v1_10072575
    • M0494L_v1_10074877
    • M0494S_v1_10072584
    • M0494L_v1_10077586
    • M0494X_v1_10077338
    • M0494S_v1_10077339
    • M0494S_v1_10080914
    • M0494L_v1_10080286
    • M0494S_v1_10082584
    • M0494X_v1_10082399
    • M0494L_v1_10083909
    • M0494S_v1_10085130
    • M0494X_v1_10086302
    • M0494L_v1_10088107
    • M0494S_v1_10088239
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    • M0494X_v1_10091664
    • M0494L_v1_10090574
    • M0494S_v1_10091798
    • M0494L_v1_10094430
    • M0494S_v1_10094427
    • M0494X_v1_10094179
    • M0494X_v1_10097057
    • M0494X_v1_10097942
    • M0494L_v1_10097776
    • M0494X_v1_10099184
    • M0494S_v1_10097777
    • M0494L_v1_10100932
    • M0494L_v1_10101679
    • M0494S_v1_10101681
    • M0494X_v1_10101246
    • M0494L_v1_10104350
    • M0494X_v1_10104713
    • M0494S_v1_10108147
    • M0494L_v1_10108305
    • M0494G_v0_10097056
    • M0494X_v1_10109831
    • M0494S_v2_10109705
    • M0494L_v2_10111768
    • M0494L_v2_10114138
    • M0494X_v2_10115009
    • M0494S_v2_10116769
    • M0494X_v2_10116382
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    • M0494L_v2_10120888
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    • M0494X_v2_10120933
    • M0494L_v2_10123780
    • M0494S_v2_10123781
    • M0494X_v2_10127955
    • M0494L_v2_10126669
    • M0494S_v2_10126662
    • M0494L_v2_10132276
    • M0494S_v2_10132512
    • M0494X_v2_10132096
    • M0494S_v2_10133637
    • M0494G_v0_10119930
    • M0494L_v2_10136909
    • M0494X_v2_10137145
    • M0494L_v2_10138598
    • M0494S_v2_10137221
    • M0494X_v2_10140519
    • M0494G_v0_10142491
    • M0494L_v2_10141212
    • M0494S_v2_10144285
    • M0494X_v2_10145964
    • M0494G_v0_10149471
    • M0494L_v2_10146691
    • M0494S_v2_10146692
    • M0494X_v2_10148907
    • M0494S_v2_10153261
    • M0494L_v2_10152072
    • M0494G_v0_10157107
    • M0494S_v2_10156983
    • M0494G_v0_10158320
    • M0494G_v0_10158514
    • M0494X_v2_10157014
    • M0494G_v0_10162775
    • M0494L_v2_10156984
    • M0494G_v1_10163600
    • M0494S_v2_10158874
    • M0494X_v2_10162269

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Q5® Hot Start High-Fidelity 2X Master Mix