DNA Polymerase Selection Chart
The following table lists properties that should be considered when choosing a polymerase. Since these properties can depend on reaction conditions, the primary references should be consulted prior to use in a given application.
| PCR Polymerases | 3′–>5′ Exonuclease | Fidelity | 5′–>3′ Exonuclease | Strand Displacement | Nick Translation | Extend RNA Primer | Extension from Nick | dU Tolerance | Resulting Ends | Popular Formats Available | Applications |
|---|---|---|---|---|---|---|---|---|---|---|---|
| High Fidelity PCR | |||||||||||
| Q5® High-Fidelity DNA Polymerase | ++++ | 280x Taq | No | _ | No | No | No | No | Blunt | Q5® High-Fidelity DNA Polymerase | Ultra high-fidelity PCR, long range PCR, cloning from in vitro material for protein expression or gene analysis, SNP analysis via cloning and sequencing, site directed mutagenesis |
| Q5® Hot Start High-Fidelity DNA Polymerase | |||||||||||
| Q5® High-Fidelity DNA Polymerase 2x Master Mix | |||||||||||
| Q5® High-Fidelity Hot Start DNA Polymerase 2x Master Mix | |||||||||||
| NEBNext® Ultra™ II Q5® Master Mix | NGS library prep | ||||||||||
| Q5®Blood Direct 2X Master Mix |
Blood direct high-fidelity PCR" |
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| Q5U® Hot Start High-Fidelity DNA Polymerase | ++++ | No | _ | No | No | No | Yes | Blunt | Q5U® Hot Start High-Fidelity DNA Polymerase | USER cloning, high fidelity amplification of bisulfite converted or deaminated DNA substrates, carryover prevention | |
| NEBNext Q5U® Master Mix | |||||||||||
| Phusion® High-Fidelity DNA Polymerase* | ++++ | 39b-50xc Taq | No | _ | No | No | No | No | Blunt | Phusion® High Fidelity DNA Polymerase | High-Fidelity PCR, cloning |
| Phusion® High Fidelity PCR Master Mix with HF Buffer | |||||||||||
| Phusion® High Fidelity PCR Master Mix with GC Buffer | |||||||||||
| Phusion® Hot Start Flex DNA Polymerase | |||||||||||
| Phusion® Hot Start Flex 2X Master Mix | |||||||||||
| Routine PCR | |||||||||||
| OneTaq® DNA Polymerase | ++ | 2x | Yes | _j | Yes | No | Yes | Yes | 3'A/Blunt | OneTaq® DNA Polymerase | Routine PCR, colony PCR, genotype screening |
| OneTaq® Hot Start DNA Polymerase | |||||||||||
| OneTaq® 2X Master Mix with Standard Buffer | |||||||||||
| OneTaq® Hot Start 2X Master Mix with Standard Buffer | |||||||||||
| OneTaq® Hot Start 2X Master Mix with GC Buffer | |||||||||||
| OneTaq® Quick-Load DNA Polymerase | |||||||||||
| OneTaq® Quick-Load® 2X Master Mix | |||||||||||
| Taq DNA Polymerase | _ |
1x (1.3-1.8x10-4) b |
Yes | _j | Yes | No | Yes | Yes | 3'A | Taq DNA Polymerase with Thermopol Buffer | Routine PCR |
| Hot Start Taq DNA Polymerase | |||||||||||
| Taq 2X Master Mix | |||||||||||
| Hot Start Taq 2X Master Mix | |||||||||||
| Quick-Load Taq 2X Master Mix | |||||||||||
| Multiplex PCR 5X Master Mix | Multiplex end point PCR | ||||||||||
| Specialty PCR | |||||||||||
| LongAmp® Taq DNA Polymerase | ++ | 2x | Yes | _j | Yes | No | Yes | Yes | 3'A/Blunt | LongAmp® Taq DNA Polymerase | Long range PCR for complex and simple templates |
| LongAmp® Hot Start Taq DNA Polymerase | |||||||||||
| LongAmp® Taq 2X Master Mix | |||||||||||
| LongAmp® Hot Start Taq 2X Master Mix | |||||||||||
| Hemo KlenTaq | _ | No | + | No | No | No | Yes | 3'A | Hemo KlenTaq | Blood direct PCR | |
| Epimark® Hot Start Taq DNA Polymerase | _ | Yes | _j | Yes | No | Yes | Yes | 3'A | Epimark® Hot Start Taq DNA Polymerase | Bisulfite converted DNA amplification, AT rich templates | |
| Isothermal Amplification and Strand Displacement | 3′–>5′ Exonuclease | Error Rate (x 10-6) |
5′–>3′ Exonuclease | Strand Displacement | Nick Translation | Extend RNA Primer | Extension from Nick | dU Tolerance | Resulting Ends | Popular Formats Available | Applications |
| Bst DNA Polymerase, Full Length | _ | Yes | _j |
Yes | Yes | Yes | Yes | 3'A | Bst DNA Polymerase, Full Length | Nick translation | |
| Bst DNA Polymerase, Large Fragment | _ | No | ++++ | No | Yes | Yes | Yes | 3'A | Bst DNA Polymerase, Large Fragment | Isothermal Amplification (LAMP, SDA) molecular diagnostic, field diagnostics | |
| Bst 2.0® DNA Polymerase | _ | 62 (±5)e | No | ++++ | No | Yes | Yes | Yes | 3'A | Bst 2.0® DNA Polymerase | |
| Bst 2.0 WarmStart® DNA Polymerase | |||||||||||
| WarmStart® Colorimetric LAMP 2X Master Mix with UDG |
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| WarmStart® LAMP Kit (DNA & RNA) | |||||||||||
| WarmStart® Colorimetric LAMP 2X Master Mix (DNA & RNA) | |||||||||||
| Bst 3.0 DNA Polymerase | _ | 70 (±23)e | No | ++++ | No | Yes | Yes | Yes | 3'A | Bst 3.0 DNA Polymerase | |
| Bsu DNA Polymerase, Large Fragment | _ | No | + | No | Yes | Yes | Yes | 3'A | Bsu DNA Polymerase, Large Fragment | Isothermal Amplification (RPA, SDA, RCA) | |
| phi29 DNA Polymerase | ++++ | 5i | No | ++++ | No | Yes | Yesk | Yes | Blunt | phi29 DNA Polymerase | RCA |
| phi29-XT DNA Polymerase | ++++ | 5i | No | ++++ | No | Yes | Yesk | Yes | Blunt | phi29-XT RCA Kit | RCA, WGA |
| phi29-XT WGA Kit |
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| Polymerases for DNA Manipulation | 3′–>5′ Exonuclease | Error Rate (x 10-6) | 5′–>3′ Exonuclease | Strand Displacement | Nick Translation | Extend RNA Primer | Extension from Nick | dU Tolerance | Resulting Ends | Popular Formats Available | Applications |
| T7 DNA Polymerase (unmodified) | ++++ | 15h | No | _ | No | Yes | No | Blunt | T7 DNA Polymerase (unmodified) | ||
| Sulfolobus DNA Polymerase IV | _ | No | _ | No | 3'A | Sulfolobus DNA Polymerase IV | Trans lesion bypass | ||||
| Therminator™ DNA Polymerase | _ | No | + | No | Yes | Yes | Yes | 3'A | Therminator™ DNA Polymerase | Chain terminator | |
| DNA Polymerase I (E. coli) | ++ | 9f | Yes | _j | Yes | Yes | Yes | Yes | Blunt | DNA Polymerase I (E. coli) | Second strand synthesis, nick translation |
| DNA Polymerase I, Large (Klenow) Fragment' | ++ | 18g | No | + | No | Yes | Yes | Yes | Blunt | DNA Polymerase I, Large (Klenow) Fragment' | Blunting, primer extension |
| Klenow Fragment (3′→5′ exo-) | _ | 100g | No | +++ | No | Yes | Yes | Yes | 3'A | Klenow Fragment (3′→5′ exo-) | A tailing, random priming labeling |
| T4 DNA Polymerase | ++++ | <1f | No | _ | No | Yes | No | Blunt | T4 DNA Polymerase | Blunting | |
| Legacy Polymerases | 3′–>5′ Exonuclease | Error Rate (x 10-6) | 5′–>3′ Exonuclease | Strand Displacement | Nick Translation | Extend RNA Primer | Extension from Nick | dU Tolerance | Resulting Ends | Popular Formats Available | Applications |
| Vent® DNA Polymerase | ++ | No | + | No | No | Yes | No | Blunt | Vent® DNA Polymerase | ||
| Vent® (exo–) DNA Polymerase | _ | No | +++ | No | No | Yes | No | 3'A | Vent® (exo–) DNA Polymerase | ||
| Deep Vent® DNA Polymerase | +++ | No | ++ | No | No | Yes | No | Blunt | Deep Vent® DNA Polymerase | ||
| Deep Vent® (exo–) DNA Polymerase | _ | No | +++ | No | No | Yes | No | 3'A | Deep Vent® (exo–) DNA Polymerase | ||
Deep Vent® and Therminator™ are trademarks of New England Biolabs, Inc.
Q5®, Q5U®, LongAmp®, OneTaq®, Vent® are registered trademarks of New England Biolabs, Inc.
* Notice to Customers:
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.
Phusion® is registered trademark and property of Thermo Fisher Scientific.
References:
- Measured by the opal reversion assay of Kunkel et al. [(1987) Proc. Natl. Acad. Sci. USA, 84, 4865–4869 PMID: 3474631] which reflects the error rate for a single round of gap_filling DNA synthesis. Several alternative assays are also available, although comparing error frequencies among these assays is complicated because they measure different aspects of error introduction.
- Potapov, V., & Ong, J. L. (2017). Examining Sources of Error in PCR by Single_Molecule Sequencing. PloS one, 12(1), e0169774. doi:10.1371/journal.pone.0169774 PMID: 28060945
- as reported my Finnzymes/Thermo Scientific
- Tindall, K.R. and Kunkel, T.A. (1988) Biochemistry, 27, 6008–6013. PMID: 2847780
- Potapov, V., Fu, X., Dai, N., Corrêa, I.R., Tanner, N.A., Ong, J.L. (2018) Base modifications affecting RNA polymerase and reverse transcriptase fidelity. Nucleic Acids Research, 46 (11), 5753–5763.doi: 10.1093/nar/gky341 PMID: 29750267
- Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984) J. Biol. Chem., 259, 1539–1545 PMID: 6229537
- Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990) J. Biol. Chem., 265, 13878–13887. PMID: 2199444
- Mattila, P., Korpela, J., Tenkanen, T. and Pitkanen, K. (1991) Nucleic Acids Res., 19, 4967–4973. PMID: 1923765
- Esteban, Salas, and Blanco 1993 JBC 268(4):2719-2726 PMID: 8428945
- Destroys displaced strand.
- Extension from a nick with Phi29 is not efficient, we would recommend Bsu or E. coli DNA Polymerase I for applications needing this activity.
- denotes no observed activity
+ denotes presence and degree of activity
