Five Quality Features of Q5
Q1 Fidelity
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)
Q2 Robustness
High specificity and yield with minimal optimization
Q3 Coverage
Superior performance for a broad range of amplicons (from high AT to high GC)
Q4 Speed
Short extension times
Q5 Amplicon length
Robust amplifications up to 20 kb for simple templates, and 10 kb for complex
Video: Important tips for Q5 Polymerase
Feature Article:
„Polymerase Fidelity –
What is it and what does it mean for your PCR?“
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.
RQ5 formats support robust amplification of a broad range of targets
PCR
Recommended products:
- Q5 High-Fidelity DNAPolymerase (NEB #M0491)
- Q5 Hot Start High-Fidelity DNA Polymerase (NEB #M0493)
- Q5U Hot Start High-Fidelity DNA Polymerase (NEB #M0515)
- Q5 High-Fidelity 2X Master Mix (NEB #M0492)
- Q5 Hot Start High-Fidelity 2X Master Mix (NEB #M0494)
- Q5 High-Fidelity PCR Kit (NEB #E0555)
- Q5 Quick-Load High-Fidelity 2X Master Mix (#M0578)
- Q5 Quick-Load Hot Start High-Fidelity 2X Master Mix (#M0580)
Available Q5 Polymerase Products
| PRODUCT | Prod. # | Size | INFO | ||
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| Q5 High-Fidelity DNA Polymerase | M0491S | 100 units |  |
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| M0491L | 500 units | |
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| Q5 High-Fidelity 2X Master Mix | M0492S | 100 rxns (50 µl/vol) | |
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| M0492L | 500 rxns (50 µl/vol) | |
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| Q5 Hot Start High-Fidelity DNA Polymerase | M0493S | 100 units | |
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| M0493L | 500 units | |
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| Q5 Hot Start High-Fidelity 2X Master Mix | M0494S | 100 rxns (2×1.25ml) | |
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| M0494L | 500 rxns (10×1.25ml) | |
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| M0494X | 500 rxns (1×12.5ml) | |
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| Q5 Quick-Load High-Fidelity 2X Master Mix | M0578L | 500 rxns | |
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| Q5 Quick-Load Hot Start High-Fidelity 2X Master Mix | M0580L | 500 rxns | |
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| Q5U Hot Start High-FidelityDNA Polymerase | M0515S | 100 units | |
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| M0515L | 500 units | |
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| Also available as Q5 High-Fidelity PCR Kit: | |||||
| Q5 High-Fidelity PCR Kit | E0555S | 50 rxns | |
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| E0555L | 200 rxns | |
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| Also available as Next-Gen-Seq optimized formulation: | |||||
| NEBNext Ultra II Q5 Master Mix | M0544S | 50 rxns (50 µl vol) | |
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| M0544L | 250 rxns (50 µl vol) | |
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| M0544X | 500 rxns (50 µl vol) | |
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| NEBNext High-Fidelity 2X PCR Master Mix | M0541S | 50 rxns (50 µl/vol) | |
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| M0541L | 250 rxns (50 µl/vol) | |
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| NEBNext Q5 Hot Start HiFi PCR Master Mix | M0543S | 50 rxns (50 µl vol) | |
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| M0543L | 250 rxns (50 µl vol) | |
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| NEBNext Q5U Master Mix | M0597S | 50 rxns (50 µl/vol) | |
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| M0597L | 250 rxns (50 µl/vol) | |
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| or for “ARTIC” workflows: | |||||
| One-Step: LunaScript Multiplex One-Step RT-PCR Kit |
E1555S | 50 rxns | |
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| E1555L | 250 rxns | |
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| Two-Step: LunaScript RT SuperMix |
M3010L | 100 rxns | |
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| M3010X | 500 rxns | |
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| M3010E | 2500 rxns | |
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| AAlso available as Direct Sample Amplification formulation: | |||||
| Q5 Blood Direct 2X Master Mix | M0500S | 100 rxns (1x1ml) | |
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| M0500L | 500 rxns (5x1ml) | |
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| Also available for Mutagenesis: | |||||
| Q5 Site-Directed Mutagenesis Kit | E0554S | 10 rxns | |
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| Q5 Site-Directed Mutagenesis Kit (Without Competent Cells) | E0552S | 10 rxns | |
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| Compatible products: | |||||
| Deoxynucleotide Solutions, Mix | N0447S | 8 µmol of each | |
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| N0447L | 40 µmol of each | |
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As of: 19.11.2025
Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.
