900,000 U of T7 DNA Ligase (3,000,000 U/mL) and 2x Rapid Ligation Buffer
The T7 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
- Only joins efficiently cohesive-end termini
- Low efficiency joining blunt ends
- Repair single-stranded nicks in duplex DNA
Product Details
T7 DNA Ligase catalyzes the formation of a phosphodiester bond between a 5ʹ phosphate and a 3ʹ hydroxyl termini in duplex DNA. The enzyme only efficiently joins cohesive end termini and repairs single-stranded nicks in duplex DNA. It is not very effective at catalyzing blunt end ligation. However, adding high concentrations of PEG 6000 at ≥ 20% (w/v) can stimulate T7 DNA Ligase activity and lead to measurable results. Generally, under normal reaction conditions, T7 DNA Ligase cannot carry out blunt-end DNA ligation, which makes it a suitable option for cases where both blunt and cohesive ends of DNA are present. Still, only cohesive ends need to be joined.
This enzyme is supplied in 20 mM Tris-HCl, 50 mM NaCl, 1 mM DTT, 0.1 mM EDTA and 50% glycerol; pH 7.5 at 25°C.
The 2x Rapid Ligation Buffer contains 132 mM Tris-HCI, 20 mM MgCl2, 2 mM DTT, 2 mM ATP and 15% PEG 6000; pH 7.6 at 25°C.
Performance
- Storage temperature: –25°C to –15°C
- Molecular weight: 41,132 Daltons
| Test | Units tested | Specification |
|---|
| Purity | n/a | >99% |
| Specific activity | n/a | 3,000,000 U/mg |
| Single-stranded exonuclease | 30,000 U | <1% released |
| Double-stranded exonuclease | 30,000 U | <1% released |
| Double-stranded endonuclease | 30,000 U | No conversion |
| E. coli DNA contamination | 30,000 U | <10 copies |