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Q5® High-Fidelity DNA Polymerase
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Catalog #
M0491S
M0491L
Size
100 units
500 units
Concentration
2,000 units/ml
2,000 units/ml
Categories: Q5® High-Fidelity DNA Polymerase Products, Q5® High-Fidelity DNA Polymerase Products
Applications: DNA Amplification, PCR and qPCR, Fast PCR, Fast PCR | More +
Product
Information
FAQs &
Tech Tips
Description
Properties and Usage
Protocols &
Manuals
Other Tools &
Resources
Quality &
Safety
Legal
Information
Advantages and Features
Related Products
Description
Q5® High-Fidelity DNA
Polymerase is a high-fidelity,
thermostable DNA polymerase
with 3´→ 5´ exonuclease
activity, fused to a processivity-enhancing Sso7d domain to support robust DNA amplification. With an error rate ~280-fold lower than that of
Taq DNA Polymerase, Q5 High-Fidelity DNA Polymerase is ideal for cloning and can be used for long or difficult amplicons. Q5 HighFidelity 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 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 High-Fidelity DNA Polymerase. All reactions
were conducted using 30 cycles of amplification and visualized by microfluidic LabChip® analysis.
Product Source
An E. coli strain that carries the Q5 High-Fidelity DNA Polymerase gene.
Reagents Supplied
The following reagents are supplied with this product:
Store at (°C)
Q5® High GC Enhancer
Q5® Reaction Buffer Pack
Concentration
5X
-20
5X
Advantages and Features
Applications
High-fidelity PCR
Cloning
Long or Difficult Amplification
High-throughput PCR
Properties and Usage
Unit Definition
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.
Storage Temperature
-20°C
Heat Inactivation
No
Unit Assay Conditions
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.
Related Products
Companion Products
Deoxynucleotide (dNTP) Solution Mix
Deoxynucleotide (dNTP) Solution Set
Magnesium Chloride (MgCl2) Solution
Q5® Hot Start High-Fidelity 2X Master Mix
Q5® High-Fidelity 2X Master Mix
Q5® Hot Start High-Fidelity DNA Polymerase
Q5® Reaction Buffer Pack
FAQs
FAQs
What are the advantages to using Q5® 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 experiment using Q5® 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?
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. There is a precipitate in the bottom of the buffer tube. Is this normal?
11. What length of product can be made by Q5® High-Fidelity DNA Polymerase?
12. I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Q5® High-Fidelity
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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?
I am competing in the iGEM competition. Do you have any products that I should consider from NEB?
Protocols
Datacards
Protocols
1. PCR Using Q5® High-Fidelity DNA Polymerase (M0491)
Datacards
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and
quality controls. The following file naming structure is used to name the majority of these document files: [Catalog
Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the
Technical Support Form for appropriate document.
M0491Datasheet-Lot0041207
M0491Datasheet-Lot0041209
M0491Datasheet-Lot0041209
M0491Datasheet-Lot0041212
M0491Datasheet-Lot0051402
M0491Datasheet-Lot0051312
M0491Datasheet-Lot0051412
M0491Datasheet-Lot0051506
Selection Charts
Usage Guidelines & Tips
Application Notes
Citations
Feature Articles
Troubleshooting Guides
NEB Publications
Selection Charts
DNA Polymerase Selection Chart
Feature Articles
Polymerase Fidelity: What is it, and what does it mean for your PCR?
Usage Guidelines & Tips
Activity of Restriction Enzymes in a Q5®, Taq or Phusion PCR Mix
Guidelines for PCR Optimization with Thermophilic DNA Polymerases
Troubleshooting Guides
PCR Troubleshooting Guide
Application Notes
Multiplex PCR using Q5 High-Fidelity DNA Polymerase
NEB Publications
Potapov Vladamir, Ong Jennifer L (2017). Examining Sources of Error in PCR by Single-Molecule Sequencing. PLoS ONE.
12(1): e0169774, PubMedID: 28060945
Citations
Yonghe Zhang, Huiming Huang, Shanshan Xu, Bo Wang, Jianhua Ju, Huarong Tan, Wenli Li (2015). Activation and
enhancement of Fredericamycin A production in deepsea-derived Streptomyces somaliensis SCSIO ZH66 by using ribosome
engineering and response surface methodology. Microb Cell Fact. 14, 64. PubMedID: 25927229, DOI: 10.1186/s12934-0150244-2
Binyamin D Berkovits, Christine Mayr (2015). Alternative 3' UTRs act as scaffolds to regulate membrane protein localization.
Nature. , PubMedID: 25896326, DOI: 10.1038/nature14321
Jun Wu, Daiji Okamura, Mo Li, Keiichiro Suzuki, Chongyuan Luo, Li Ma, Yupeng He, Zhongwei Li, Chris Benner, Isao Tamura,
Marie N Krause, Joseph R Nery, Tingting Du, Zhuzhu Zhang, Tomoaki Hishida, Yuta Takahashi, Emi Aizawa, Na Young Kim,
Jeronimo Lajara, Pedro Guillen, Josep M Campistol, Concepcion Rodriguez Esteban, Pablo J Ross, Alan Saghatelian, Bing
Ren, Joseph R Ecker, Juan Carlos Izpisua Belmonte (2015). An alternative pluripotent state confers interspecies chimaeric
competency. Nature. , PubMedID: 25945737, DOI: 10.1038/nature14413
Longhai Dai, Can Liu, Yueming Zhu, Jiangsheng Zhang, Yan Men, Zeng Yan, Yuanxia Sun (2015). Functional Characterization
of Cucurbitadienol Synthase and Triterpene Glycosyltransferase Involved in Biosynthesis of Mogrosides from Siraitia grosvenorii.
Plant Cell Physiol. , PubMedID: 25759326, DOI: 10.1093/pcp/pcv043
Silva-Herzog E, McDonald EM, Crooks AL, Detweiler CS. (2015). Physiologic Stresses Reveal a Salmonella Persister State and
TA Family Toxins Modulate Tolerance to These Stresses PLoS One. 12, PubMedID: 26633172, DOI: 10.1371
Amin Zargar, David N Quan, Milad Emamian, Chen Yu Tsao, Hsuan-Chen Wu, Chelsea R Virgile, William E Bentley (2015).
Rational design of 'controller cells' to manipulate protein and phenotype expression. Metab Eng. , PubMedID: 25908186, DOI:
10.1016/j.ymben.2015.04.001
Christine Henke, Pamela L Strissel, Maria-Theresa Schubert, Megan Mitchell, Claus C Stolt, Florian Faschingbauer, Matthias
W Beckmann, Reiner Strick (2015). Selective expression of sense and antisense transcripts of the sushi-ichi-related
retrotransposon - derived family during mouse placentogenesis. Retrovirology. 12, 9. PubMedID: 25888968, DOI:
10.1186/s12977-015-0138-8
Harish Nag Kankipati, Marta Rubio-Texeira, Dries Castermans, George Diallinas, Johan M Thevelein (2015). Sul1 and Sul2
Sulfate Transceptors Signal to Protein Kinase A upon Exit of Sulfur Starvation. J Biol Chem. 290, 10430-46. PubMedID:
25724649, DOI: 10.1074/jbc.M114.629022
Yuan Xue, Jossef Osborn, Anand Panchal, Jay L Mellies (2015). The RpoE Stress Response Pathway Mediates Reduction of
the Virulence of Enteropathogenic Escherichia coli by Zinc. Appl Environ Microbiol. 81, 3766-74. PubMedID: 25819956, DOI:
10.1128/AEM.00507-15
Yang YJ, Han YY, Chen K, Zhang Y, Liu X, Li S, Wang KQ, Ge JB, Liu W, Zuo J. (2015). TonEBP modulates the protective
effect of taurine in ischemia-induced cytotoxicity in cardiomyocytes Cell Death Dis.. PubMedID: 26673669, DOI: 10.1038
Yafeng Li, Delu Song, Ying Song, Liangliang Zhao, Natalie Wolkow, John W Tobias, Wenchao Song, Joshua L Dunaief (2015).
Iron-induced Local Complement Component 3 (C3) Up-regulation via Non-canonical Transforming Growth Factor (TGF)-β
Signaling in the Retinal Pigment Epithelium. J Biol Chem. 290, 11918-34. PubMedID: 25802332, DOI: 10.1074/jbc.M115.645903
Wang XJ, Zhang XJ, Hu W, Zhang TY, Wang SQ (2014). A simple and efficient strategy for the de novo construction of greaterthan-genome-length hepatitis B virus replicons J Virol Methods. 207, 158-62. PubMedID: 25025817, DOI:
10.1016/j.jviromet.2014.07.009
Bert De Rybel, Milad Adibi, Alice S. Breda, Jos R. Wendrich, Margot E. Smit, Ondej Novk, Nobutoshi Yamaguchi, Saiko
Yoshida, Gert Van Isterdael, Joakim Palovaara, Bart Nijsse, Mark V. Boekschoten, Guido Hooiveld, Tom Beeckman, Doris
Wagner, Karin Ljung, Christian Fleck, Dolf Weijers (2014). Integration of growth and patterning during vascular tissue formation
in Arabidopsis Science. 345, 1255215. PubMedID: 25104393, DOI: 10.1126/science.1255215
Vidhyadhar Nandana, Sushant Singh, Abhay Narayan Singh, Vikash Kumar Dubey (2014). Procerain B, a cysteine protease
from Calotropis procera, requires N-terminus pro-region for activity: cDNA cloning and expression with pro-sequence. Protein
Expr Purif. 103C, 16-22. PubMedID: 25173974, DOI: 10.1016/j.pep.2014.08.003
Martin Kostovcik, Craig C Bateman, Miroslav Kolarik, Lukasz L Stelinski, Bjarte H Jordal, Jiri Hulcr (2014). The ambrosia
symbiosis is specific in some species and promiscuous in others: evidence from community pyrosequencing. ISME J. ,
PubMedID: 25083930, DOI: 10.1038/ismej.2014.115
Xin Duan, Arjun Krishnaswamy, Irina De la Huerta, Joshua R Sanes (2014). Type II Cadherins Guide Assembly of a DirectionSelective Retinal Circuit. Cell. 158, 793-807. PubMedID: 25126785, DOI: 10.1016/j.cell.2014.06.047
Wilber Quispe-Tintaya, Ryan R White, Vasily N Popov, Jan Vijg, Alexander Y Maslov (2013). Fast mitochondrial DNA isolation
from mammalian cells for next-generation sequencing Biotechniques. 55(3), 133-6. PubMedID: 24003945, DOI:
10.2144/000114077
Hicham Bouabe, Klaus Okkenhaug (2013). A Protocol for Construction of Gene Targeting Vectors and Generation of
Homologous Recombinant Embryonic Stem Cells Methods in Molecular Biology. 1064, 337-354. PubMedID: 23996269
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: 23266826
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. PubMedID: 23907213
Quality Control
Certificate of Analysis
Specifications
Datacards
Safety Data Sheet
Quality Control
Quality Control Assays
The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product.
Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting
Documents section of this page. Further information regarding NEB product quality can be found here.
Endonuclease Activity (Nicking):
The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to
the nicked form is determined by agarose gel electrophoresis.
PCR Amplification (Enhancer Dependent, GC-rich):
The polymerase is tested in a polymerase chain reaction (PCR) using a GC-rich control template and specific primers, resulting
in the enhancer-dependent production of the expected product.
PCR Amplification (Master Mix):
The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers,
resulting in the expected product.
Protein Purity (SDS-PAGE):
The physical purity is assessed by comparing contaminating protein bands in a concentrated sample to the protein of interest
band in a sample of known dilution. The purity is determined by SDS-PAGE.
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's
for an individual lot. The following file naming structure is used to name these document files: [Product
Number]_[Size]_[Version]_[Lot Number]
M0491S_L_v2_0051506
M0491S_L_v2_0051512
M0491S_L_v2_0051606
M0491S_L_v2_0051612
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the
product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
M0491S_L_v2
Safety Data Sheet
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
Q5® High-Fidelity DNA Polymerase
Q5® High GC Enhancer
Q5® Reaction Buffer Pack
Datacards
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and
quality controls. The following file naming structure is used to name the majority of these document files: [Catalog
Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the
Technical Support Form for appropriate document.
M0491Datasheet-Lot0041207
M0491Datasheet-Lot0041209
M0491Datasheet-Lot0041209
M0491Datasheet-Lot0041212
M0491Datasheet-Lot0051402
M0491Datasheet-Lot0051312
M0491Datasheet-Lot0051412
M0491Datasheet-Lot0051506
Legal and Disclaimers
Legal and Disclaimers
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc
(NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain
additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes
in humans or animals.
Licenses
This product is covered by one or more Patents.
This product is licensed from Bio-Rad Laboratories, Inc. under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645 and
corresponding patents in other countries for use only in: (a) standard (non-real time) PCR in the research field only, but not realtime PCR or digital PCR; (b) any in-vitro diagnostics application, except for applications using real-time or digital PCR; and (c) any
non-PCR applications in DNA sequencing, isothermal amplification and the production of synthetic DNA.
Trademarks
Q5® is a registered trademark of New England Biolabs, Inc.