A new standard for High Fidelity PCR
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, even lower than Phusion Polymerase
Engineered for Performance
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.
Q5 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.
Video: Q5 High-Fidelity Polymerase
Q5 DNA Polymerase is also available as a hot start version. The Q5 Hot Start DNA Polymerase utilizes NEBs aptamer technology, allowing room temperature reaction setup without the need of an additional activation step.
Five Quality Features of Q5
The highest fidelity amplification available (~280X higher than Taq, ~7x higher than Phusion Polymerase)”Ultra-Fidelity” (~280X higher than Taq, ~7x higher than Phusion Polymerase)
High specificity and yield with minimal optimization
Superior performance for a broad range of amplicons (from high AT to high GC)
Short extension times
Q5 Amplicon length
Robust amplifications up to 20 kb for simple templates, and 10 kb for complex
NEU: Q5U Hot Start High-Fidelity DNA Polymerase
Q5U is a modified version of Q5 High-Fidelity DNA Polymerase containing a mutation in the uracil-binding pocket that enables the ability to read and amplify templates containing uracil and inosine bases.
This is useful for amplifying bisulfite-converted, enzymatically-deaminated, or damaged DNA, preventing carryover contamination in PCR (when used with dUTP and UDG), and in USER cloning methods.
Optimized protocols are available for recommended applications.
Comparison of High-Fidelity Polymerases
1 We continue to investigate improved assays to characterize Q5’s very low error rate to ensure thatwe 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)
3 Template not specified.
4 Simple templates include plasmid, viral and E. coli genomic DNA. Complex templates include plant, human and other mammalian genomic DNA.
5 Values provided by individual manufacturers.
Amplification Performance Across a Wide Range of Genomic Targets
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.
Available Q5 Polymerase Products
As of: 01.01.2020