T4 DNA ligase
Cat.No. : | CMC12 |
Availability | March 29, 2025 |
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Source : | E.coli |
Tag : | Non |
Description : | T4 DNA Ligase is a recombinant enzyme formulated and tested for high activity in blunt- and cohesive-end ligations for cloning and linker addition. The enzyme covalently joins 5"-phosphate and 3"-hydroxyl termini in duplex DNA or RNA. It is free of detectable exo- or endonuclease activities. |
Applications : | Cloning of restriction fragments; Joining linkers and adapters to blunt-ended DNA. |
Unit Definition (Cohesive End Ligation Unit) : | One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of lambda DNA (5" DNA termini concentration of 0.12μM, 300μg/ml) in a total reaction volume of 20μl in 30 minutes at 16°C in 1× T4 DNA Ligase Reaction Buffer. One Cohesive End Ligation Unit equals 0.015 Weiss (ATP-PP exchange) units, or one Weiss unit equals 67 Cohesive End Ligation Units. |
Concentration : | 400,000 units/ml. |
Quality Control Tests : | Activity, SDS-PAGE (purity), DNase, endonuclease/nickase, RNase. |
Storage : | T4 DNA ligase in 10mM Tris-HCl, 50mM KCl, 1mM dithiothreitol, 0.1mM EDTA, 0.2 mg/ml BSA, 50% glycerol, pH 7.4 (25℃) should be stored at -20°C. |
Heat Inactivation : | 65°C for 10 minutes. |
1× T4 DNA Ligase Reaction Buffer : | 50mM Tris-HCl, 10mM MgCl2, 1mM ATP, 10mM dithiothreitol, 25μg/ml BSA, pH7.5 (25℃)). |
Reaction Conditions : | 1× T4 DNA Ligase Reaction Buffer, incubate at 16°C. For convenience, ligations may be done at room temperature (20-25°C). For cohesive (sticky) ends, use 1μl of T4 DNA Ligase in a 20μl reaction for 10 minutes. For blunt ends, use 1μl of T4 DNA Ligase in a 20μl reaction for 2 hours. |
◆ Recombinant Proteins | ||
CMC12 | T4 DNA ligase | +Inquiry |
Not For Human Consumption!
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