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Taq DNA Ligase

$50.00$195.00


    2,000 units (50 µl) - $50.00 (Cat.# 3218 )

    10,000 units (250 µl) - $195.00 (Cat.# 3219 )

Category: .

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. Taq DNA Ligase 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).

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Product Source
E. coli strain expressing the cloned Taq DNA ligase gene from Thermus aqauticus 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 Ligase 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 Ligase 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

Taq DNA Ligase is free from detectable RNase or contaminating DNA endonuclease activities.

Additional information

Units

2,000 units (50 µl), 10,000 units (250 µl)

  1. Set-up the reaction as follows:

Protocol

 

 

 

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.

Taq DNA Ligase

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