Product Description

Taq DNA Ligase catalyzes the formation of a phosphodiester bond in duplex DNA containing adjacent 5′-phosphoryl and 3′-hydroxyl termini, using NAD as a cofactor. The ligation will occur only if the oligonucleotides are perfectly paired to the complementary target DNA and have no gaps between them; therefore, a single-base substitution can be detected. This product is active at elevated temperatures (45°C-70°C) (1, 2).

The physical purity of this enzyme is ≥99% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).

Product Source
E. coli strain expressing the cloned Taq DNA ligase gene from Thermus aquaticus HB8

Applications

• Allele-specific gene detection by using Ligase Detection Reaction (LDR) and Ligase Chain Reaction (LCR) ⁽¹⁾.
• Mutagenesis by incorporation of a phosphorylated oligonucleotide during primer extension amplification⁽³⁾.

Product Includes
1)   Taq DNA Ligase
2)   10X Taq DNA Ligase Buffer with NAD  

Storage Temperature
–20 °C

Storage Buffer
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA,
50% Glycerol, pH 7.5 @ 25 ºC

10X Taq DNA reaction buffer with NAD

500 mM Tris-HCl, 100 mM MgCl₂, 100 mM DTT, 10 mM NAD , pH 7.5 @ 25ºC

Unit Definition

One unit is defined as the amount of Taq DNA required to join 50% of 1 μg of the 12-base cohesive ends of Lambda DNA cut with Sma I and Sal I in 50 μl reaction in 15 min incubation at 45 °C.

Quality Control

This product is free from detectable RNase or contaminating DNA endonuclease activities.

 

1. Set-up the reaction as follows:

2. Incubate at 50 °C for 15-30 minutes.

References
1. Barany, F. (1991). Proc. Natl. Acad. Sci. USA. 88, 189-193.
2. Takahashi, M. et al. (1984). J. Biol. Chem. 259, 10041-10047.
3. Michael, S.F. (1994). Biotechniques. 16, 411-412.

 

 

Manual – Taq DNA Ligase
MSDS – IG Taq DNA Ligase

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