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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
Want to try this product? Request a sample Need a custom/large volume order? Contact Us
Bulk packaging may also be available and requested for large recurring orders. Learn More
产品信息
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-High Fidelity 2X Master Mix formats allow robust amplification of a broad range of targets with a single formulation.
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.
产品来源
An E. coli strain that carries the Q5 High-Fidelity DNA Polymerase gene.
产品类别:
Q5® High-Fidelity DNA Polymerases Products,
Master Mixes Products
应用:
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,
PCR,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB
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
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
热失活
否
优势和特性
应用特性
High-specificity PCR
High-fidelity 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
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
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.
Product specifications for individual components in Q5 Hot Start High-Fidelity 2X Master Mix are available separately.
参考文献
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
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,
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
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
What are the advantages to using Q5® Hot Start High-Fidelity 2X Master Mix?
What is the fidelity of Q5® High-Fidelity DNA Polymerase?
How should I determine an appropriate annealing temperature for my reaction?
What should my primer concentration be when using Q5® High-Fidelity DNA Polymerase products?
How should I set up a PCR reaction using Q5® Hot Start 2X Master Mix?
My template is GC rich or supercoiled. How can I optimize my product yield using Q5® High-Fidelity Master Mixes?
There is a precipitate in the bottom of the Master Mix tube? Is this normal?
How do I activate Q5® Hot Start High-Fidelity DNA Polymerase?
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?
What length of product can be made by Q5® High-Fidelity DNA Polymerase?
I have a tube of the Q5® High GC Enhancer from another product formulation – can I add it to the Q5 Master Mix?
What are the stability and storage requirements of the Q5® Master Mixes?
Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
Where can I find help troubleshooting my PCR?
I’d like to clone a fragment amplified with Q5® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
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
产品引用文献查找工具
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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
Want to try this product? Request a sample 
Need a custom/large volume order? Contact Us 
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