For details on NEB's quality controls for DNA ligases, visit our Ligase Quality page.
A salt tolerant variant of T4 DNA Ligase, Salt-T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA or RNA and is designed to function at higher salt concentrations than wild type T4 DNA Ligase. This enzyme will join cohesive end termini at salt concentrations as high as 300 mM without any loss in activity. This enzyme is insensitive to salt carried over from other reaction components (vector or insert DNA) and allows ligation reactions to proceed in alternative reaction buffers with higher levels of salt (e.g., NEBuffer 3.1).
Figure 1: Salt-T4 DNA Ligase displays increased salt tolerance on a cohesive end substrate as compared to T4 DNA Ligase.
400 units of T4 DNA Ligase (NEB #M0202) or Salt-T4 DNA Ligase (NEB #M0467) was incubated with 0.12 μM HindIII (cohesive end) fluorescent-labeled fragments at 25°C for 10 minutes in the presence of 1X T4 DNA Ligase Buffer containing increasing amounts of salt (0-1000 mM NaCl). Ligation products were detected by capillary electrophoresis.
T4 DNA Ligase rapidly loses activity at salt concentrations above 200 mM, whereas Salt-T4 DNA Ligase retains > 60% activity on a cohesive end substrate at salt concentrations as high as 500 mM.
Product Source
An E. coli strain that carries a plasmid encoding the engineered Salt-T4 DNA Ligase gene.
This product is related to the following categories:
One unit is defined as the amount of enzyme required to give 50% ligation of 6 µg of Lambda-HindIII DNA in 30 minutes at 25°C in a total reaction volume of 20 µ in 1X T4 DNA Ligase Reaction Buffer supplemented with 100 mM NaCl.
Reaction Conditions
1X T4 DNA Ligase Reaction Buffer
Incubate at 25°C
T4 DNA Ligase Reaction Buffer
50 mM Tris-HCl
10 mM MgCl2
1 mM ATP
10 mM DTT
(pH 7.5 @ 25°C)
Storage Buffer
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
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