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Home >Technical Resource > Polymerases > Properties of DNA 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

Vent_R

Vent_R (exo-)

Deep
Vent_R

Deep
Vent_R
(exo-)

9°N_m

Ther-
minator

T7
DNA

E. coli
Poly I

Klenow
Frag
Poly. I

Klenow
Frag
exo-

M-MulV
Reverse
Trans-
criptase

T4
DNA

f29

Phu-
sion

DyNA
zyme
EXT

DyNA
zyme
II HS

5'—>3' Exonuclease

3'—>5' Exonuclease

+++

++++

Error Rate^a (x10^-6)

285^c

57^b

190^b

15^b

9^q

18^w

100^w

<1^q

.44

Strand Displacement

++++

++^g

+++^g

+++^x

+++

+++++

Nick Translation

Thermal Stability

+++

Km dNTPs

13 µM^g

60 µM^g

40 µM^g

50 µM^g

80 µM^x

18 µM^s

1-2 µM^h

2 µM^k

18 µM^p

2 µM^u

0.5µM^z

Km DNA^d

2 nM^g

0.1 nM^g

0.01 nM^g

0.05 nM^x

18 nM^s

5 nM^h

Extend RNA Primer^y

Extension From Nick

References:
a. 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.
b. Mattila, P., Korpela, J., Tenkanen, T. and Pitkanen, K. (1991) Nucleic Acids Res. 19, 4967–4973.
c. Tindall, K.R. and Kunkel, T.A. (1988) Biochemistry 27, 6008–6013.
d. Km values for DNA are expressed in terms of moles of primer-template complexes.
e. Joyce, C.M. (1989) J. Biol. Chem. 264, 10858–10866.
g. Kong, H.M., Kucera, R.B. and Jack, W.E., J. Biol. Chem. (1993) 268, 1965–1975.
h. McClure, W.R. and Jovin, T.M. (1975) J. Biol. Chem. 250, 4073–4080.
k. Polesky, A.H., Steitz, T.A., Grindley, N.D.F. and Joyce, C.M. (1990) J. Biol. Chem. 265, 14579–14591.
n. Tabor, S., Huber, H.E. and Richardson, C.C. (1987) J. Biol. Chem. 262, 16212–16223.
o. Tabor, S. and Richardson, C.C. (1987) Proc. Natl. Acad. Sci. USA 84, 4767–4771.
p. Ricchetti, M. and Buc, H. (1990) EMBO J. 9, 1583–1593.
q. Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984) J. Biol. Chem. 259, 1539–1545.
s. Patel, S.S., Wong, E. and Johnson, K.A. (1991) Biochemistry 30, 511–525.
u. Gillin, F.D. and Nossal, N.G. (1975) Biochem. Biophys. Res. Commun. 64, 457–464.
v. Cline, J., Braman, J.C. and Hogrefe, H.H. (1996) Nucleic Acids Res. 24, 3546–3551.
w. Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990) J. Biol. Chem. 265, 13878–13887.
x. Southworth, M.W. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 5281–5285.
y. 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).
z. Saturno, J., Blanco, L., Salas, M. and Esteban, J.A. (1995) J. Biol. Chem. 270, 31235–31243.