Q5 High Fidelity DNA Polymerase - New England Biolabs France (2024)

Fidelity at its finest

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 Bufferprovides 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.

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)
³ Takagi et al (1997) Appl. Env. Microbiol. 63, 4504-4510.

Details

Figure 1: Robust amplification with Q5 (A) and Q5 Hot Start (B) High-Fidelity DNA Polymerases, regardless of GC content

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.

Figure 2: 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.

Figure 3: Q5 DNA Polymerase offers superior amplification for a wide range of templates, regardless of GC content

Amplification of a variety of human genomic amplicons from low to high GC content demonstrates the broad performance of Q5 High-Fidelity DNA Polymerase. All reactions were conducted using 20 ng of input template and included 30 cycles of amplification. Results were visualized by microfluidic LabChip® analysis. Competitor polymerases were cycled according to manufacturer’s recommendations. For the final three amplicons, GC Buffers or enhancers were used when supplied with the polymerase.

Figure 4:Q5 Hot Start DNA Polymerase offers superior amplification for a wide range of templates, regardless of GC content Amplification

Amplification of a variety of human genomic amplicons from low to high GC content demonstrates the broad performance of Q5 Hot Start High-Fidelity DNA Polymerase. All reactions were set up at room temperature using 20 ng of input template and included 30 amplification cycles. Results were visualized by microfluidic LabChip® analysis. Competitor polymerases were cycled according to manufacturer’s recommendations. For the final three amplicons, GC Buffers or enhancers were used when provided with the polymerase.

Table: Comparison ofHighFidelity DNA Polymerases

¹ PCR-based mutation screening in lacZ (NEB), lacI (Agilent) or rpsL (Life)
² Takagi et al (1997) Appl. Env. Microbiol. 63, 4504-4510.
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.

Available Products

Phusion® is a registered trademark and property of Thermo Fisher Scientific. Phusion® DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific. AccuPrime™ is a trademark of Life Technologies, Inc. Platinum® is a registered trademark of Life Technologies, Inc. PfuUltra™ is a trademark of Agilent Technologies, Inc. Q5®, Deep Vent™ and 9°N™ are trademarks of New England Biolabs, Inc. Vent™ is a registered trademark of New England Biolabs, Inc. LabChip® is a registered trademark of Caliper Life Sciences, part of Perkin Elmer, Inc.