Q5 High-Fidelity DNA Polymerase

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.

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¹ We continue to investigate improved assays to characterize Q5’s very low error rate to ensure that we present the most accurate fidelity data possible (Potapov, V. and Ong, J.L. (2017) PLoS ONE. 12(1): e0169774).
² PCR-based mutation screening in lacZ (NEB), lacI (Agilent) or rpsL (Life).


Check out the specifications and application data of the different formats of this high-fidelity enzyme:

Q5 High-Fidelity DNA Polymerase

Q5 High-Fidelity 2X Master Mix

Q5 Hot Start High-Fidelity DNA Polymerase

Q5 Hot Start High-Fidelity 2X Master Mix

Advantages

• 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.

Amplification of a variety of human genomic amplicons from low to high GC content using either Q5 or Q5 Hot Start High-Fidelity DNA Polymerase. Reactions using Q5 Hot Start were set up at room temperature. All reactions were conducted using 30 cycles of amplification and visualized by microfluidic LabChip® analysis.

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.

PCR was performed with a variety of amplicons, with GC content ranging from high AT to high GC, with Q5 and several other commercially available polymerases. All polymerases were cycled according to manufacturer's recommendations, including use of GC Buffers and enhancers when recommended. Yield and purity of reaction products were quantitated and represented, as shown in the figure key, by dot color and size. A large dark green dot represents the most successful performance. Q5 provides superior performance across the range of GC content.

Master Mix and Stand-Alone Formats Provide Convenience and Flexibility

Why Choose Q5 High-fidelity 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".

Important Tips for Q5 Polymerase

Here are some quick tips for getting the most out of NEB's Q5® High-Fidelity DNA Polymerase.

Q5® High-Fidelity DNA Polymerases Feedback

"I have started using Q5 for single cell PCR and it is working brilliantly. I get very bright bands and am able to derive sequences from nearly all the positive wells."

Postdoc, King’s College London

"My Q5 sample arrived just as I was struggling with what I had anticipated would be a routine Phusion reaction. After a couple of failures with Phusion, I tried the Q5 sample, head-to-head, under a few different reaction conditions. All of my Q5 reactions worked beautifully, while all of the Phusion reactions failed. After that observation, I repeated a recent successful Phusion reaction with both Phusion and Q5 to compare the products in circumstances under which Phusion worked very well. In that set of tests, all reactions were successful, but the crude gels showed more product in each Q5 reaction and clearer, cleaner bands. Since Q5 is priced competitively compared to Phusion, we have started replacing Phusion with Q5 in our standard reactions. So far, we have not seen any reactions work with Phusion and not with Q5. We hope this will continue to be the case! I have started recommending Q5 to other scientists who are faced with tricky PCR reactions. Two months ago, I would've recommended Phusion, but now it's Q5."

Lab Manager, Colorado State University

"Q5 High Fidelity DNA Polymerase from NEB is a superb product. I am currently using this to amplify some of my DNA inserts and have repeatedly come up with very positive results better than what I got using another brand not in any way associated with NEB. Am I surprised by the result I am getting from Q5? Absolutely not! Any bioscience kit with NEB on it must be good."

Researcher, University of North Texas

"Q5® works great. It was able to amplify a very difficult product, one I honestly didn't think would work. I am extremely happy with the Q5 enzyme."

Research Asst., Vanderbilt University

"We switched from KOD to phusion polymerase which at the time seemed the best. But now, the new Q5 polymerase is among the best available in the market, even better than phusion. Highly recommended to others!"

Ph.D. Student, University of Cambridge

"100% satisfaction: very specific and vast synthesis of the PCR product on BAC-clone template."

Research Associate, Fordham University

"Not only did I observe great amplification of my product but there were no contaminating bands and the desired amplicon was as tight as any I have ever seen."

Research Assistant, Colorado State University

"We work on an organism with a high GC content and the gene I was amplifying is 70% GC. The 1.9kb band was gorgeous and I got nothing from Phusion. I love it. It will be our new lab polymerase."

Professor, Washington University School of Medicine

"Q5 DNA Polymerase gave me wonderful results on the first shot."

Research technologist, University of Nebraska Medical Center

"The Q5 works great, way better than the Pfu Turbo I was using. I have already ordered Q5 polymerase for my experiments."

Postdoc, MIT

"I have a 75% GC rich region that I need to amplify and tried all possible polymerases one can think of. Nothing worked. With Q5 I was so happily surprised when I looked at the gel and saw single bands of the right size!"

Customer, NIBR

Sample request Q5 High-Fidelity DNA Polymerase

See what our customers are saying about Q5® High-Fidelity DNA Polymerases. Do you want to test yourself? Fill in your details below and receive a Q5 DNA Polymerase sample**.

**This free sample can be requested by laboratory from the Netherlands, Belgium and Luxembourg . No cash or cash equivalent. Offer may not be applied to existing, pending or prior orders. Free samples cannot be requests by current users of the product.