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Q5® Hot Start High-Fidelity DNA Polymerase
Fidelity at its finest
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)
Hot start and master mix formats available
精选视频
Important Tips for Q5® Polymerase
观看其他视频
Interested in glycerol-free or custom formulations? – contact our Custom Solutions Group
Bulk packaging may also be available and requested for large recurring orders. Learn More
产品信息
Q5® Hot Start High-Fidelity DNA Polymerase is 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 DNA Polymerase is ideal for cloning and can be used for long or difficult amplicons. Q5 Hot Start High-Fidelity DNA Polymerase is supplied with an optimized buffer system that allows robust amplification regardless of GC content. The 5X Q5 Reaction Buffer contains 2 mM Mg++ at final (1X) reaction concentrations and is recommended for most routine applications. For GC-rich targets (≥ 65% GC), amplification can be improved by the addition of the 5X Q5 High GC Enhancer. Q5 Hot Start High-Fidelity DNA Polymerase 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.
Amplification of a variety of human genomic amplicons from low to high GC content using Q5 Hot Start High-Fidelity DNA Polymerase. 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
应用:
Genome Editing Applications,
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
M0493S
-20
Q5® Hot Start High-Fidelity DNA Polymerase
M0493SVIAL
-20
1 x 0.05 ml
2,000 units/ml
Q5® Reaction Buffer Pack
B9027SVIAL
-20
1 x 1.5 ml
5 X
Q5® High GC Enhancer
B9028AVIAL
-20
1 x 1.5 ml
5 X
M0493L
-20
Q5® Hot Start High-Fidelity DNA Polymerase
M0493LVIAL
-20
1 x 0.25 ml
2,000 units/ml
Q5® Reaction Buffer Pack
B9027SVIAL
-20
4 x 1.5 ml
5 X
Q5® High GC Enhancer
B9028AVIAL
-20
4 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.
热失活
否
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
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 2X Master Mix
Q5® High-Fidelity DNA Polymerase
Q5® Hot Start High-Fidelity 2X Master Mix
Q5® Reaction Buffer Pack
操作说明、说明书 & 用法
操作说明
PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)
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 DNA Polymerase?
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 High-Fidelity DNA Polymerase?
My template is GC rich or supercoiled. How can I optimize my product yield using Q5® High-Fidelity DNA Polymerase?
Do I need to modify my annealing temperature when using the Q5® High GC Enhancer?
When should I add the High GC Enhancer?
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?
There is a precipitate in the bottom of the buffer tube. Is this normal?
What length of product can be made by Q5® High-Fidelity DNA Polymerase?
I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Q5® High-Fidelity DNA Polymerase?
Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
Where can I find help troubleshooting my PCR?
Will Q5® High-Fidelity DNA Polymerase incorporate dUTPs?
I’d like to clone a fragment amplified with Q5® High-Fidelity DNA Polymerase. Do I have to blunt-end clone?
Do other polymerases work in Q5® Reaction Buffer?
What is the difference between Q5 and Q5U Hot Start High-Fidelity DNA Polymerase?
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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更多引用文献
Stoczynska-Fidelus E, Och W, Rieske P, Bienkowski M, Banaszczyk M, Winiecka-Klimek M, Zieba J, Janik K, Rosiak K, Treda C, Stawski R, Radomiak-Zaluska A, Piaskowski S (2014) Spontaneous in vitro senescence of glioma cells confirmed by an antibody against IDH1R132H Anticancer Res; 34(6), 2859-67. PubMedID: 24922649
Currin A, Swainston N, Day PJ, Kell DB (2014) SpeedyGenes: an improved gene synthesis method for the efficient production of error-corrected, synthetic protein libraries for directed evolution Protein Eng Des Sel; 27(9), 273-80. PubMedID: 25108914, DOI: 10.1093/protein/gzu029
Juliane Krebes, Richard D. Morgan, Boyke Bunk, Cathrin Spröer, Khai Luong, Raphael Parusel, Brian P. Anton, Christoph König, Christine Josenhans, Jörg Overmann, Richard J. Roberts, Jonas Korlach, Sebastian Suerbaum (2013) The complex methylome of the human gastric pathogen Helicobacter pylori Nucleic Acids Res; 42(4), 2415-2432. PubMedID: 24302578, DOI: 10.1093/nar/gkt1201
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