Q5® Hot Start High-Fidelity DNA Polymerase (2024)

HomeProductsQ5^® Hot Start High-Fidelity DNA Polymerase

• Product Information

  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 T_m Calculator is highly recommended.

  

  Product Source

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

  This product is related to the following categories:

  Q5® High-Fidelity DNA Polymerases Products

  This product can be used in the following applications:

  Hot Start PCR,

  Long Range PCR,

  Fast PCR,

  Genome Editing Applications,

  High-Fidelity PCR,

  Multiplex PCR,

  Specialty PCR,

  Routine PCR,

  PCR,

  Fast Cloning: Accelerate your cloning workflows with reagents from NEB

  • Reagents Supplied

    The following reagents are supplied with this product:

    NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
    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

  • See Also

    NEB Interactive Tools - New England Biolabs GmbHPCR Using Q5® High-Fidelity DNA Polymerase (M0491)NEB - PCR Using Q5® High-Fidelity DNA Polymerase (M0491)NEB Interactive Tools - New England Biolabs GmbH

    Properties & 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.

    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.

  • Advantages and Features

    Application Features

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

  • Related Products

    Companion Products

    • 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

• Protocols, Manuals & Usage

  • Protocols

    1. PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)

  • Usage Guidelines

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

  • Application Notes

    • AppNote_Multiplex_PCR_Using_Q5_HF_DNA_Polymerase

• Tools & Resources

  • Selection Charts

    • DNA Polymerase Selection Chart

  • Web Tools

    • PCR Selector
    • Tm Calculator

• FAQs & Troubleshooting

  • FAQs

    1. What are the advantages of using Q5® High-Fidelity DNA Polymerase?
    2. What are the differences between the numerous Q5® Polymerase products available?
    3. My results are not as expected. Where can I find troubleshooting help?
    4. What ends will my PCR products have?
    5. What does exonuclease activity mean for a DNA polymerase?
    6. Does Q5® High-Fidelity DNA Polymerase exhibit a strand displacement activity?
    7. What are the properties of this polymerase (fidelity, product ends, max amplicon, modified base incorporation, etc.)?
    8. What should the final primer concentration be in my PCR?
    9. When and how should I use the Q5® High GC Enhancer?
    10. Do other polymerases work in Q5® Reaction Buffer?
    11. There is a precipitate in the bottom of the Master Mix tube? Is this normal?
    12. What are Hot Start and WarmStart® polymerases and when would I use them?

    See Also

    NEB Interactive Tools - New England Biolabs GmbH

  • Troubleshooting

    • PCR Troubleshooting Guide

• Citations & Technical Literature

  • Citations

    Product Citation Tool

    Powered by Bioz See more details on Bioz

    Additional Citations

    • 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
    • Toru Takahashi, Ken Maeda, Tadaki Suzuki, Aki Ishido, Toru Shigeoka, Takayuki Tominaga, Toshiaki Kamei, Masahiro Honda, Daisuke Ninomiya, Takenori Sakai, Takanori Senba, Shozo Kaneyuki, Shota Sakaguchi, Akira Satoh, Takanori Hosokawa, Yojiro Kawabe, Shintaro Kurihara, Koichi Izumikawa, Shigeru Kohno, Taichi Azuma, Koichiro Suemori, Masaki Yasukawa, Tetsuya Mizutani, Tsutomu Omatsu, Yukie Katayama, Masaharu Miyahara, Masahito Ijuin, Kazuko Doi, Masaru Okuda, Kazunori Umeki, Tomoya Saito, Kazuko f*ckushima, Kensuke Nakajima, Tomoki Yoshikawa, Hideki Tani, Shuetsu f*ckushi, Aiko f*ckuma, Momoko Ogata, Masayuki Shimojima, Noriko Nakajima, Noriyo Nagata, Harutaka Katano, Hitomi f*ckumoto, Yuko Sato, Hideki Hasegawa, Takuya Yamagishi, Kazunori Oishi, Ichiro Kurane, Shigeru Morikawa, Masayuki Saijo (2013) The First Identification and Retrospective Study of Severe Fever With Thrombocytopenia Syndrome in Japan J Inf Dis; 209(6), 816-827. PubMedID: 24231186

  • Publications

    • 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

  • Feature Articles

    • Using aptamers to control enzyme activities: Hot Start Taq and beyond

• Quality, Safety & Legal

  • Quality Control Assays

    Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

  • Specifications & Change Notifications

    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]

    • M0493S_L_v1

  • Certificate of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    • M0493S_L_v1_0121606
    • M0493S_L_v1_0071505
    • M0493S_L_v1_0111512
    • M0493S_L_v1_0151612
    • M0493S_L_v1_0151706
    • M0493S_L_v1_0191712
    • M0493L_v1_10007543
    • M0493L_v1_10014733
    • M0493S_v1_10017673
    • M0493L_v1_10021411
    • M0493S_v1_10022245
    • M0493L_v1_10025839
    • M0493S_v1_10026171
    • M0493L_v1_10032192
    • M0493S_v1_10029216
    • M0493L_v1_10034496
    • M0493S_v1_10036747
    • M0493L_v1_10036748
    • M0493L_v1_10042498
    • M0493S_v1_10040297
    • M0493L_v1_10045790
    • M0493L_v1_10047950
    • M0493S_v1_10047831
    • M0493L_v1_10049885
    • M0493L_v1_10051820
    • M0493S_v1_10051814
    • M0493L_v1_10055338
    • M0493S_v1_10057910
    • M0493L_v1_10057912
    • M0493L_v1_10060871
    • M0493S_v1_10060286
    • M0493L_v1_10063522
    • M0493L_v1_10065197
    • M0493S_v1_10067254
    • M0493L_v1_10071140
    • M0493L_v1_10075123
    • M0493L_v1_10077342
    • M0493S_v1_10077190
    • M0493S_v1_10078997
    • M0493L_v1_10079277
    • M0493L_v1_10082201
    • M0493L_v1_10085116
    • M0493S_v1_10083166
    • M0493L_v1_10088131
    • M0493L_v1_10089130
    • M0493S_v1_10088152
    • M0493S_v1_10093297
    • M0493L_v1_10093298
    • M0493L_v1_10098032
    • M0493S_v1_10098036
    • M0493L_v1_10100657
    • M0493S_v1_10100656
    • M0493L_v1_10102587
    • M0493L_v1_10106979
    • M0493S_v1_10103356
    • M0493L_v1_10109421
    • M0493L_v1_10111051
    • M0493L_v1_10112649
    • M0493S_v1_10110303
    • M0493S_v1_10114447
    • M0493L_v1_10114919
    • M0493S_v1_10119830
    • M0493L_v1_10118912
    • M0493L_v1_10122244
    • M0493L_v1_10127960
    • M0493S_v1_10127959
    • M0493L_v1_10131148
    • M0493L_v1_10134956
    • M0493L_v1_10138439
    • M0493S_v1_10140760
    • M0493S_v1_10145147
    • M0493L_v1_10144277
    • M0493L_v1_10148895
    • M0493L_v1_10151327
    • M0493S_v1_10153091
    • M0493L_v1_10155132
    • M0493S_v1_10157140
    • M0493L_v1_10162258
    • M0493S_v1_10164060
    • M0493L_v1_10166985
    • M0493S_v1_10170232
    • M0493L_v1_10172094
    • M0493L_v1_10176590
    • M0493S_v1_10174968
    • M0493L_v1_10179239
    • M0493S_v1_10180974
    • M0493L_v1_10183701
    • M0493L_v1_10191817
    • M0493S_v1_10191819
    • M0493S_v1_10202495
    • M0493L_v1_10206180
    • M0493S_v1_10208638
    • M0493S_v1_10218685
    • M0493L_v1_10219588
    • M0493L_v1_10224871
    • M0493L_v1_10212821
    • M0493L_v1_10229462
    • M0493S_v1_10226659
    • M0493L_v1_10236856

  • Safety Data Sheets

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    • Q5® Hot Start High-Fidelity DNA Polymerase

    • Q5® Reaction Buffer Pack

    • Q5® High GC Enhancer

  • Legal and Disclaimers

    Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

    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.

    New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

    Licenses

    This product is covered by one or more patents.

    This product is licensed for research and commercial use 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. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows.

    For additional information or to inquire about commercial use, please contact busdev@neb.com.

    Trademarks

    NEW ENGLAND BIOLABS^®and Q5^®are registered trademarks of New England Biolabs, Inc.

    LABCHIP^®is a registered trademark of Caliper Life Sciences, Inc., part of PerkinElmer, Inc.
    SYBR^®is a registered trademark of Molecular Probes, Inc., now owned by Life Technologies, Inc.