Taq DNA ligase
Cat. No.: SB-G3459-2000U
Size: 2000U
Description
Taq DNA ligase is derived from Thermus thermophilus HB8 ligase and is recombinantly expressed in Escherichia coli. It is a thermostable DNA ligase that can catalyze the formation of phosphate diphosphate between the 5'-phosphate and 3'-hydroxyl of two adjacent DNA strands. ester bond. Taq DNA Ligase uses NAD as a cofactor, which is active at temperatures from 37°C–75°C.
- Source : derived from Thermus thermophilus HB8 is recombinantly expressed in Escherichia coli.
- Definition of enzyme activity: 1 enzyme is defined as the amount of enzyme required to ligate 50% of 1 μg of BstEII-digested λDNA fragments (12 bp sticky ends) in a 50 μL reaction system at 45°C for 15 minutes living unit.
Purity and concentration: SDS-PAGE detection purity ≥ 95%; endogenous nucleic acid residue < 1 pg/μL (qPCR detection); 40 U/μL.
- Enzyme Storage Buffer: 10 mM Tris-HCl, 50 mM KCL, 1 mM DTT, 0.1 mM EDTA, 200 µg/ml BSA, 50% Glycerol, pH 7.4.
- 10x Taq Reaction Buffer: 200 mM Tris-HCl, 250 mM Potassium Acetate, 100 mM Magnesium Acetate, 10 mM NAD+, 100 mM DTT, 1% Triton® X-100, pH 7.6.
Storage and transportation
Transport in wet ice packs; store at -20°C, valid for 12 months.
Product Contents
| Component Number |
Component |
G3459 |
| G3459-1 |
Taq DNA ligase |
50 μL |
| G3459-2 |
10х Taq Reaction buffer |
500 μL |
| Product Manual |
1 copy |
Steps
1. Configure the reaction system according to the following table:
| Component |
Volume |
| DNA sample |
1 μg |
| 10х Taq Reaction Buffer |
5 μL |
| Taq DNA ligase |
2 μL |
| Nuclease Free Water |
To 50 μL |
2. Incubate the reaction system at 45°C for 15 minutes. After the reaction was complete, 50 mM EDTA was added to terminate the reaction.
Precautions
1. 10×Taq DNA Ligase Buffer contains the coenzyme factor NAD + , in order to prolong the half-life of NAD + , the reaction buffer should be stored at -80°C;
2. For your safety and health, please wear a lab coat and disposable gloves for operation.
