M.B. Enzymes GmbH

ThermolasePR - High-fidelity DNA polymerase with proofreading activity.

Description

ThermolasePR (EC 2.7.7.7) is a thermostable DNA-dependent DNA polymerase cloned from the archaea Thermococcus litoralis and overexpressed in recombinant E.coli strain. The enzyme has 5´-3´ polymerase, 5´-3´ exonuclease (strand displacement) as well as 3´-5´ exonuclease (”proofreading“) activities. The fidelity of ThermolasePR DNA polymerse is 10-fold greater as that of Taq DNA polymerase (error rate is 4.5-5.7x10^-5). The enzyme is extremely thermostable (half-life at 95°C 6.7 h, temperature optimum is 70-75°C). Up to 15 kb can be amplified from lambda DNA and up to 10 kb from genomic DNA.

Unit definition

One unit of the enzyme will incorporate 10 nmol of dNTPs into an acid-insoluble form in 30 min at 72°C under standard assay conditions using a DNA template.

Reaction buffer

B1 (x10, with Mg²⁺ ): 670 mM Tris-HCl, pH 8.8, 166 mM (NH₄)_₂SO₄, 15 mM MgCl₂, 0.1% Tween 20.

Storage

Store at -20°C.

Applications

high-fidelity PCR; long template and accurate PCR.

Quality control

activity, SDS-PAGE purity, specific performance tests.

References

1. Mattila, P. et al. (1991) Nucleic Acids Res. 19, 4967-4973. 2. Kong, H.M. et al. (1993) J. Biol. Chem 268, 1965-1975. 3. Perler, F. et al. (1992) Proc. Natl. Acad. Sci. USA 89, 5577. 4. Belkin, S., Jannasch, H.W. (1985) Arch. Microbiol. 141, 181-186.

Catalog #

T8.2

T8.3

Pack size

250 U

500 U

ThermolasePR - High-fidelity DNA polymerase with proofreading activity.
Description	ThermolasePR (EC 2.7.7.7) is a thermostable DNA-dependent DNA polymerase cloned from the archaea Thermococcus litoralis and overexpressed in recombinant E.coli strain. The enzyme has 5´-3´ polymerase, 5´-3´ exonuclease (strand displacement) as well as 3´-5´ exonuclease (”proofreading“) activities. The fidelity of ThermolasePR DNA polymerse is 10-fold greater as that of Taq DNA polymerase (error rate is 4.5-5.7x10^-5). The enzyme is extremely thermostable (half-life at 95°C 6.7 h, temperature optimum is 70-75°C). Up to 15 kb can be amplified from lambda DNA and up to 10 kb from genomic DNA.
Unit definition	One unit of the enzyme will incorporate 10 nmol of dNTPs into an acid-insoluble form in 30 min at 72°C under standard assay conditions using a DNA template.
Reaction buffer	B1 (x10, with Mg²⁺ ): 670 mM Tris-HCl, pH 8.8, 166 mM (NH₄)_₂SO₄, 15 mM MgCl₂, 0.1% Tween 20.
Storage	Store at -20°C.
Applications	high-fidelity PCR; long template and accurate PCR.
Quality control	activity, SDS-PAGE purity, specific performance tests.
References	1. Mattila, P. et al. (1991) Nucleic Acids Res. 19, 4967-4973. 2. Kong, H.M. et al. (1993) J. Biol. Chem 268, 1965-1975. 3. Perler, F. et al. (1992) Proc. Natl. Acad. Sci. USA 89, 5577. 4. Belkin, S., Jannasch, H.W. (1985) Arch. Microbiol. 141, 181-186.
Catalog #	T8.2	T8.3
Pack size	250 U	500 U