Fidelity at its Finest.
Q5® High-Fidelity DNA Polymerase (NEB #M0491) 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.
- 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
The Q5 buffer system is designed to provide superior performance with minimal optimization across a broad range of amplicons, regardless of GC content. For routine or complex amplicons up to ~65% GC content, Q5 Reaction Buffer (NEB #B9027) provides reliable and robust amplification. For amplicons with high GC content (>65% GC), addition of the Q5 High GC Enhancer ensures continued maximum performance. Q5 and Q5 Hot Start DNA Polymerases are available as standalone enzymes, or in a master mix format for added convenience. Master mix formulations include dNTPs, Mg++ and all necessary buffer components.
In contrast to chemically modified or antibody-based hot start polymerases, NEB's Q5 Hot Start (NEB #M0493) utilizes a unique synthetic aptamer. This molecule binds to the polymerase through non-covalent interactions, blocking activity during the reaction setup. The polymerase is activated during normal cycling conditions, allowing reactions to be set up at room temperature. Q5 Hot Start does not require a separate high temperature activation step, shortening reaction times and increasing ease-of-use. Q5 Hot Start Polymerase is an ideal choice for high specificity amplification and provides robust amplification of a wide variety of amplicons, regardless of GC content.
Q5® is a registered trademark of New England Biolabs, Inc.
LabChip® is a registered trademark of Caliper Life Sciences, part of Perkin Elmer, Inc.
- PCR Using Q5® High-Fidelity DNA Polymerase (M0491)
- Protocol for a Routine PCR (E0555)
- PCR Optimization (E0555)
- PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)
- Protocol for Q5® Hot Start High-Fidelity 2X Master Mix
- Protocol for Q5® High-Fidelity 2X Master Mix
- PCR Using NEBNext® High-Fidelity 2X PCR Master Mix (M0541)
- Protocol for a PCR reaction using NEBNext® Q5® Hot Start HiFi PCR Master Mix (M0543)
- Protocol for a PCR reaction using NEBNext® Ultra™ II Q5® Master Mix (M0544)
Anatomy of a Polymerase - How Structure Effects Function
Understanding Variability in DNA Amplification Reactions
The PCR brochure provides product information on the wide range of DNA polymerases available from NEB, including tools for selection and troubleshooting tips.
- DNA Polymerase Selection Chart
- PCR Troubleshooting Guide
- Guidelines for PCR Optimization with Thermophilic DNA Polymerases
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- Fidelity – the highest fidelity amplification available (>100X higher than Taq)
- Robustness – high specificity and yield with minimal optimization
- Coverage – superior performance for a broad range of amplicons (from high AT to high GC)
- Speed – short extension times
- Amplicon length – robust amplifications up to 20 kb for simple templates, and 10 kb for complex
Template/product specificity: Is RNA or DNA involved? Is the 3´ terminus at a gap, nick or at the end of the template?
Removal of existing nucleotides: Will the nucleotide(s) be removed from the existing polynucleotide chain as part of the protocol? If so, will they be removed from the 5´ or the 3´ end?
Thermal stability: Does the polymerase need to survive incubation at high temperature or is heat inactivation desirable?
Fidelity: Will subsequent sequence analysis or expression depend on the fidelity of the synthesized products?
Here are some quick tips for getting the most out of NEB's Q5® High-Fidelity DNA Polymerase.
Not sure why Q5® is your best choice for high-fidelity amplification of GC-rich targets? NEB's scientists will show you why we call Q5 an "ultra-high fidelity polymerase".
Looking for tips on dealing with GC-bias in DNA amplification? NEB scientists have the expertise you need!
Make sure you're using the optimal polymerase for your DNA amplifications. Get tips on choosing the right DNA Polymerase for your application.