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Taq DNA Ligase
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Molecular Biological Enzymes
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Product description
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Product Description
Taq DNA Ligase Taq DNA Ligase is a high-temperature ligase that can catalyze the formation of phosphodiester between the 5'-phosphate and 3'-hydroxyl groups of two adjacent oligonucleotide strands that hybridize with the same complementary target DNA strand. key. The catalytic reaction only occurs when the two oligonucleotide strands are completely 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 substitutions. Taq DNA ligase uses NAD+ as a coenzyme factor and is active in the range of 45°C to 65°C.
Source: Recombinant Escherichia coli.
Specification: 40 U/µL.
Applications
1. Use ligase detection reaction and ligase chain reaction for specific detection of alleles;
2. Incorporation of phosphorylated oligonucleotides through PCR amplification for mutagenesis;
3. Homologous recombination.
Transportation and storage methods
Ice pack transportation. Store the product at -20°C.
Activity definition
In a 50 μL reaction system, incubating at 45°C for 15 min can make 50% of 1 μg of BstEII digested lambda DNA fragments (12 bp sticky ends) ligated. The amount of enzyme required is defined as 1 activity unit (U) .
Quality Inspection
Endonuclease activity: In a 50 μL reaction system, 10 μL enzyme solution and 0.5 μg pENZuC DNA were incubated at 37°C for 4 h. After agarose gel electrophoresis, there was no visible circular cut DNA.
Exonuclease activity: In a 50 μL reaction system, 10 μL enzyme solution and 10000 cpm/μg radiolabeled DNA are incubated at 37°C for 4 hours, and the release of soluble TCA in the solution is less than 5.0%.
Purity
SDS-PAGE: ≥99%.
Operation Manual
1. Reaction system (50 μL):
Taq DNA Ligase Taq DNA Ligase is a high-temperature ligase that can catalyze the formation of phosphodiester between the 5'-phosphate and 3'-hydroxyl groups of two adjacent oligonucleotide strands that hybridize with the same complementary target DNA strand. key. The catalytic reaction only occurs when the two oligonucleotide strands are completely 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 substitutions. Taq DNA ligase uses NAD+ as a coenzyme factor and is active in the range of 45°C to 65°C.
Source: Recombinant Escherichia coli.
Specification: 40 U/µL.
Applications
1. Use ligase detection reaction and ligase chain reaction for specific detection of alleles;
2. Incorporation of phosphorylated oligonucleotides through PCR amplification for mutagenesis;
3. Homologous recombination.
Transportation and storage methods
Ice pack transportation. Store the product at -20°C.
Activity definition
In a 50 μL reaction system, incubating at 45°C for 15 min can make 50% of 1 μg of BstEII digested lambda DNA fragments (12 bp sticky ends) ligated. The amount of enzyme required is defined as 1 activity unit (U) .
Quality Inspection
Endonuclease activity: In a 50 μL reaction system, 10 μL enzyme solution and 0.5 μg pENZuC DNA were incubated at 37°C for 4 h. After agarose gel electrophoresis, there was no visible circular cut DNA.
Exonuclease activity: In a 50 μL reaction system, 10 μL enzyme solution and 10000 cpm/μg radiolabeled DNA are incubated at 37°C for 4 hours, and the release of soluble TCA in the solution is less than 5.0%.
Purity
SDS-PAGE: ≥99%.
Operation Manual
1. Reaction system (50 μL):
| Reagent | Sample volume |
| DNA | up to 1 µg |
| 10×Taq DNA Ligase Buffer | 5μL |
| Taq DNA Ligase(40 U/µL) | 2μL |
| ddH2O | up to 50 μL |
2. Reaction conditions: incubate at 45°C for 15 min. Add stop dye solution (50% glycerol, 50 mM EDTA and bromophenol blue) to stop the reaction.
Precautions
1. The 10×Taq DNA Ligase Buffer contains the coenzyme factor NAD+. In order to prolong the half-life of NAD+, the buffer should be stored at -80℃;
2. For your safety and health, please wear lab coats and disposable gloves for operation.
Precautions
1. The 10×Taq DNA Ligase Buffer contains the coenzyme factor NAD+. In order to prolong the half-life of NAD+, the buffer should be stored at -80℃;
2. For your safety and health, please wear lab coats and disposable gloves for operation.
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