M.B. Enzymes GmbH

Tth DNA ligase -Thermostable DNA ligase.

Description

Tth DNA ligase (EC 6.5.1.2) catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA structures that are stable at high temperatures. The enzyme is stable at elevated temperatures (half-life at 65°C is 48 h, and at 95°C 30 min). The enzyme will not join ss nucleic acids, blunt-ends and has no activity on dsRNA and RNA:DNA hybrids.

Unit definition

1 unit catalyzes the ligation of 50% of the cos sites in 1 µg lambda in 1 min at 45°C using standart reaction conditions.

Reaction buffer

B18 (x10): 200 mM Tris-HCl pH 8.3, 250 mM KCl, 100 mM MgC₂, 100 mM DTT, 10 mM NAD, 1% Triton X-100.

Reaction conditions

A typical 20 ml LCR reaction requires 5U of the enzyme. Optimum temperature is 24-37°C for ligating cohesive termini and 65-72°C for closing nicks.

Storage

Store at -20°C to +4°C at high concentration (>500U/ml).

Applications

mutation analysis (ligase chain reaction (LCR), repeat expansion detection (RED);, ligation under high-stringency hybridization conditions.

Quality control

activity, SDS-PAGE/purity, absence of RNase and DNase activities, specific performance tests.

References

1. Schalling, M. et al. (1993) Nat. Genetics 4, 135. 2. Nakazawa, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 360. 3. Sirugo, G., Kidd, K. (1995) Epicentre Forum 2, 1.

Catalog #

T13.3

T13.4

Pack size

500 U

1.000 U

Tth DNA ligase -Thermostable DNA ligase.
Description	Tth DNA ligase (EC 6.5.1.2) catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA structures that are stable at high temperatures. The enzyme is stable at elevated temperatures (half-life at 65°C is 48 h, and at 95°C 30 min). The enzyme will not join ss nucleic acids, blunt-ends and has no activity on dsRNA and RNA:DNA hybrids.
Unit definition	1 unit catalyzes the ligation of 50% of the cos sites in 1 µg lambda in 1 min at 45°C using standart reaction conditions.
Reaction buffer	B18 (x10): 200 mM Tris-HCl pH 8.3, 250 mM KCl, 100 mM MgC₂, 100 mM DTT, 10 mM NAD, 1% Triton X-100.
Reaction conditions	A typical 20 ml LCR reaction requires 5U of the enzyme. Optimum temperature is 24-37°C for ligating cohesive termini and 65-72°C for closing nicks.
Storage	Store at -20°C to +4°C at high concentration (>500U/ml).
Applications	mutation analysis (ligase chain reaction (LCR), repeat expansion detection (RED);, ligation under high-stringency hybridization conditions.
Quality control	activity, SDS-PAGE/purity, absence of RNase and DNase activities, specific performance tests.
References	1. Schalling, M. et al. (1993) Nat. Genetics 4, 135. 2. Nakazawa, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 360. 3. Sirugo, G., Kidd, K. (1995) Epicentre Forum 2, 1.
Catalog #		T13.3	T13.4
Pack size		500 U	1.000 U