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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
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® and OneTaq® Hot Start 2x Master Mixes 
OneTaq® Quick-Load DNA Polymerase  
OneTaq® Quick-Load® 2X Master Mix 
Taq DNA Polymerase _ 1x Yes _j Yes No Yes Yes 3'A Taq DNA Polymerase with Thermopol Buffer Routine PCR
Hot Start TaqDNA Polymerase 
Taq and 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  _i 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® 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 WGA
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:

  1. 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. 
  2. 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
  3. as reported my Finnzymes/Thermo Scientific
  4. Tindall, K.R. and Kunkel, T.A. (1988) Biochemistry, 27, 6008–6013. PMID: 2847780
  5. 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
  6. Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984) J. Biol. Chem., 259, 1539–1545 PIMD: 26229537
  7. Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990) J. Biol. Chem., 265, 13878–13887. PIMD: 2199444
  8. Mattila, P., Korpela, J., Tenkanen, T. and Pitkanen, K. (1991) Nucleic Acids Res., 19, 4967–4973. PMID: 8420970
  9. Esteban, Salas, and Blanco 1993 JBC 268(4):2719_2726
  10. Destroys displaced strand.
  11. 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

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