PCR Selection Tool
For over 35 years, New England Biolabs, Inc. has been a world leader in the discovery and production of reagents for the life science industry. NEB offers a wide range of DNA polymerases and, through our commitment to research, ensures the development of innovative, high-quality tools for PCR.
When choosing a polymerase for PCR, we recommend starting with OneTaq® (NEB #M0480) or Q5® DNA Polymerase (NEB #M0491) for routine PCR applications, and NEBNext® High-Fidelity Master Mix (NEB #M0541) for next gen sequencing (NGS) library construction (shown below in orange). These all offer robust amplification, and can be used on a wide range of templates (routine, AT- and GC-rich). Q5 and NEBNext Master Mix (a Q5 formulation) provide the added benefit of maximum fidelity.
★ indicates recommended choice for application
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STANDARD PCR | HIGH-FIDELITY PCR | SPECIALTY PCR | |||||
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OneTaq Hot Start |
Fidelity |
Amplicons |
Sequencing |
PCR |
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Phusion(2) Flex |
LongAmp Hot Start Taq |
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| PROPERTIES | ||||||||
| Fidelity vs. Taq | 2X | 1X | ~280X(1) | >50X | 5-6X | 2X | 1X | ND |
| Amplicon Size | <6 kb | ≤5 kb | ≤20 kb | ≤20 kb | ≤6 kb | ≤30 kb | ≤1 kb | ≤2 kb |
| Extension Time | 1 kb/min | 1 kb/min | 6 kb/min | 4 kb/min | 1 kb/min | 1.2 kb/min | 1 kb/min | 0.5 kb/min |
| Resulting Ends | 3' A/Blunt | 3' A | Blunt | Blunt | Blunt | 3' A/Blunt | 3' A | 3' A |
| 3'→5' exo | Yes | No | Yes | Yes | Yes | Yes | No | No |
| 5'→3' exo | Yes | Yes | No | No | No | Yes | Yes | No |
| Units/50µl Reaction | 1.25 | 1.25 | 1.0 | 1.0 | 0.5-1.0 | 5.0 | 1.25 | N/A |
| Annealing Temperature | Tm-5 | Tm-5 | Tm+3 | Tm+3 | Tm-5 | Tm-5 | Tm-5 | Tm-5 |
| APPLICATIONS | ||||||||
| Routine PCR | ★ | ● | ● | ● | ● | ● | |
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| Colony PCR | ★ |
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| Enhanced Fidelity |
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★ | ● | ● | ● | |
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| High Fidelity | |
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★ | ● | |
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| High Yield | ★ | ● | ★ | ● | |
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| Fast | |
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★ | ● | |
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| Long Amplicon | |
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★ | ● | |
★ | |
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| GC-rich Targets | ★ | |
★ | |
● | ● | |
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| AT-rich Targets | ★ | ● | ★ | ● | |
● | ★ | |
| High Throughput | ● | ● | ● | ● | |
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| Multiplex PCR | ● | ★(3) | ● | ● | |
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| Extraction-free PCR | |
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★ |
| DNA-labeling | |
★ | |
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| Site-directed Mutagenesis | |
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★ | ● | |
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| Bisulfite Sequencing | |
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★ | |
| NGS APPLICATIONS | ||||||||
| NGS Library Amplification | ● | |
★(4) | ● | |
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| FORMATS | ||||||||
| Hot Start Available | ● | ● | ● | ● | |
● | ● | |
| Kit | |
● | ● | ● | |
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| Master Mix Available | ● | ● | ● | ● | |
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| Direct Gel Loading Available | ● | ● | |
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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) Phusion® DNA Polymerase was developed by Finnzymes Oy, now a part of Thermo Fisher Scientific.
This product is manufactured by New England Biolabs, Inc. under agreement with, and under the performance specifications of Thermo Fisher Scientific.
(3) Use Multiplex PCR 5X Master Mix.
(4) Use NEBNext High-Fidelity 2X PCR Master Mix.
Hemo KlenTaq™ is a trademark of DNA Polymerase Technology, Inc.
OneTaq®, Q5®, NEBNext®, Vent® and EpiMark® are registered trademarks of New England Biolabs, Inc.
Deep Vent® is a trademark of New England Biolabs, Inc.
Phusion® is a registered trademark of Thermo Fisher Scientific.
