T4 DNA Ligase, Recombinant
465.58 $
For reserch purpose only
Shipping cost plus VAT not included , delivery in 7-14 business days
Catalog No
USB-209228
Supplier
US Biologicals
Size
20KU
Product Overview
Product Properties
Target Gene
Product Overview
| Description | T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′ -phosphate and 3′-hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Source: Recombinant protein corresponding to T4 DNA Ligase, expressed in E. coli lambda lysogen NM 989. Unit Definition: 1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of lamda DNA (5’ DNA termini concentration of 0.12uM, 300ug/ml) in a total reaction volume of 20ul in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer. 2. One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C. Activity: One Weiss unit is equivalent to ~67 cohesive-end ligation units. • T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is >200mM. • Ligation of blunt-ended and single-base pair overhang fragments requires about 50X as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 (10% w/v final concentration), hexamine chloride or by reducing the ATP concentration to 50uM. Heat Inactivation: T4 DNA Ligase can be inactivated by incubation at 65°C for 10 minutes. T4 DNA Ligase Reaction Buffer: 50mM Tris-HCl, pH 7.8 at 25°C, 10mM magnesium chloride, 10mM DTT, 1mM ATP, 25ug/ml BSA. Reaction Conditions: 1X T4 DNA Ligase Reaction Buffer and DNA (0.1-1uM in 5´ termini). Incubate at 16°C. Applications: Suitable for use in Cloning of restriction fragments, and joining linkers and adapters to blunt-ended DNA. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| Source Antigen | Recombinant, E. coli lambda lysogen NM 989 |
| Reactivity | |
| Cross Reactivity | |
| Applications | |
| Molecular Weight |
Product Properties
| Storage | -20°C |
| Grade | Purified |
| Purity | Purified |
| Form | Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 50mM potassium chloride, 0.1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Reactivity Life | 12 months |
| Note | For reserch purpose only |
Target Gene
NA
