Product Description

Taq DNA Ligase is a thermostable ligase that catalyzes the formation of phosphodiester bonds between the 5´-phosphate and 3´-hydroxyl groups of two adjacent oligonucleotide strands that hybridize to the same complementary target DNA strand. This catalytic reaction occurs only when the two oligonucleotide strands are perfectly paired with the complementary target DNA, and there is no gap between the two oligonucleotide strands. Therefore, it can be used to detect single base substitution. Taq DNA ligase uses NAD+ as a coenzyme factor and is active at 45°C-65°C.

Product Application

1. Allele-specific detection by ligase detection reaction and ligase chain reaction;

2. Incorporating phosphorylated oligonucleotides for mutagenesis by PCR amplification;

3. Homologous recombination.

Unit Definition

One unit(U) was defined as the amount of enzyme required to ligate 50% of 1 μg of BstEII-digested λ DNA fragments (12 bp cohesive ends) after 15 min incubation at 45°C, in a 50 μL reaction system.

Quality Control

Detection of exonuclease residues: 40 U of this product and 0.5 μg of λDNA-Hind III were incubated at 37°C for 4 h, and the electrophoretic bands of DNA did not change.

Detection of nickase residues: 40 U of this product and 0.5 μg of IL23R plasmid were incubated at 37 °C for 4 h, and the electrophoretic bands of DNA did not change.

Shipping and Storage

The product is shipped with ice packs and can be stored at -20°C for one year. Please avoid repeated freeze-thaw. For long-term storage (more than 30 days), the buffer should be stored at -80°C.

Cautions

1. 10×Taq DNA Ligase Buffer contains coenzyme factor NAD+, in order to prolong the half-life of NAD+, the Buffer should be stored at -80°C.

2. Taq DNA Ligase cannot replace T4 DNA ligase.

3. For your safety and health, please wear lab coats and disposable gloves for operation.

4. This product is for research use ONLY！