2500 U of E. coli DNA Ligase (10,000 U/mL) and 10x E. coli DNA Ligase Reaction Buffer
The E. coli DNA Ligase is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Features
- Ligates DNA at nicks and cohesive termini
- Requires NAD+
Product Details
E. coli DNA ligase catalyzes the formation of phosphodiester bonds between an adjacent 5ʹ phosphate and a 3ʹ hydroxyl of DNA ends, requiring NAD+ and Mg2+ as cofactors. The ligation of blunt-ended DNA is extremely inefficient relative to cohesive DNA end ligation and nick sealing.
This enzyme is supplied in 10 mM Tris-HCl, 50 mM KCl. 1 mM DTT, 0.1 mM EDTA and 50% glycerol; pH 7.5 at 25°C.
The E. coli DNA Ligase Reaction Buffer with a concentration of 10x includes 300 mM Tris-HCl, 40 mM MgCl2, 260 µM NAD, 10 mM DTT, and 0.5 mg/mL BSA at pH 8.0 and a temperature of 25°C.
Performance
- Storage temperature: –25°C to –15°C
- Molecular weight: 73,606 Daltons
| Test | Units tested | Specification |
|---|
| Purity | n/a | >99% |
| Specific activity | n/a | 20,000–28,880 U/mg |
| Single-stranded exonuclease | 100 U | <2% released |
| Double-stranded exonuclease | 100 U | <1% released |
| Double-stranded endonuclease | 100 U | No conversion |
| E. coli DNA contamination | 50 U | <10 copies |