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Home >  Technical Reference > Polymerases
Properties of DNA Polymerases
A wide variety of polymerases have been characterized and made available for use in molecular biology. All polymerases add to the 3´ OH end of a primer in a template directed reaction. Several factors govern which polymerase should be used in a given application, including:
Template/product specificity: Is RNA or DNA involved? Is the 3´ terminus at a gap, nick or at the end of the template?
Removal of existing nucleotides: Will nucleotide(s) be removed from the existing polynucleotide chain as part of the protocol? If so, will they be removed from the 5´ or the 3´ end?
Thermal stability: Does the polymerase need to survive incubation at high temperature or is heat inactivation desirable?
Fidelity: Will subsequent sequence analysis or expression depend on the fidelity of the second strand synthesis?
The following table lists properties which should be considered in the choice of polymerases. Since these properties can depend on reaction conditions, the primary references should be consulted prior to use in a given application. For more information on the listed uses, please consult the reference lists accompanying the individual products.
 
  Bst
 DNA
L. Frag		 Taq VentR VentR
 (exo-)		 Deep
VentR		 Deep
VentR
(exo-)		 9°Nm Ther-
mina-tor		 Ther-
mina-tor II 		T7
DNA		 E. coli
Poly I		 Klenow
Frag
Poly. I		 Klenow
Frag
exo-		 M-MulV
Reverse
Trans-
criptase		 T4
DNA		 phi29 Phu-
sion		 Phu-
sion
HS		 DyNA
zyme
EXT		 DyNA
zyme
II HS
5'—>3' Exonuclease - + - - - - - - - - + - - - - - - - + +
3'—>5' Exonuclease - - ++ - +++ - + - - ++++ ++ ++ - - ++++ ++++ ++++ ++++ + -
Error Ratea (x10-6)   285c 57b 190b           15b 9q 18w 100w   <1q   .44 .44    
Strand Displacement ++++ - ++g +++g ++ ++ +++x + + - - ++ ++ +++ - +++++ - - + -
Nick Translation - + - - - - - - - - + - - - - - - - + +
Thermal Stability + ++ +++ +++ +++ +++ +++ +++ +++ - - - - - - - +++ +++ +++ ++
Km dNTPs   13 µMg 60 µMg 40 µMg 50 µMg   80 µMx     18 µMs 1-2 µMh 2 µMk   18 µMp 2 µMu 0.5µMz        
Km DNAd   2 nMg 0.1 nMg 0.1 nMg 0.01 nMg   0.05 nMx     18 nMs 5 nMh                  
Extend RNA Primery + - - - - - - + + + + + +   + + - - - -
Extension From Nick + + + + + + + + + - + + +   - + + + + +


References:

  1. Measured by the opal reversion assay of Kunkel et al. [(1987) Proc. Natl. Acad. Sci. USA, 84, 4865–4869] 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. Mattila, P., Korpela, J., Tenkanen, T. and Pitkanen, K. (1991) Nucleic Acids Res., 19, 4967–4973.
  3. Tindall, K.R. and Kunkel, T.A. (1988) Biochemistry, 27, 6008–6013.
  4. Km values for DNA are expressed in terms of moles of primer-template complexes.
  5. Joyce, C.M. (1989) J. Biol. Chem., 264, 10858–10866.
  6. Kong, H.M., Kucera, R.B. and Jack, W.E., (1993) J. Biol. Chem., 268, 1965–1975.
  7. McClure, W.R. and Jovin, T.M. (1975) J. Biol. Chem., 250, 4073–4080.
  8. Polesky, A.H., Steitz, T.A., Grindley, N.D.F. and Joyce, C.M. (1990) J. Biol. Chem., 265, 14579–14591.
  9. Tabor, S., Huber, H.E. and Richardson, C.C. (1987) J. Biol. Chem., 262, 16212–16223.
  10. Tabor, S. and Richardson, C.C. (1987) Proc. Natl. Acad. Sci. USA, 84, 4767–4771.
  11. Ricchetti, M. and Buc, H. (1990) EMBO J., 9, 1583–1593.
  12. Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984) J. Biol. Chem., 259, 1539–1545.
  13. Patel, S.S., Wong, E. and Johnson, K.A. (1991) Biochemistry, 30, 511–525.
  14. Gillin, F.D. and Nossal, N.G. (1975) Biochem. Biophys. Res. Commun., 64, 457–464.
  15. Cline, J., Braman, J.C. and Hogrefe, H.H. (1996) Nucleic Acids Res., 24, 3546–3551.
  16. Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990) J. Biol. Chem., 265, 13878–13887.
  17. Southworth, M.W. et al. (1996) Proc. Natl. Acad. Sci. USA, 93, 5281–5285.
  18. Incorporation of dNTPs was compared using a single-stranded M13 DNA template with either RNA or DNA primers (L. Greenough and W.E. Jack, unpublished observations).
  19. Saturno, J., Blanco, L., Salas, M. and Esteban, J.A. (1995) J. Biol. Chem., 270, 31235–31243.
  20. Destroys displaced strand.