T4 DNA

T4 DNA Ligase 55.3kDa protein 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.

E.Coli, cloned gene-30, bacteriophage T4.

400U/ul solution.

50% glycerol, 20mM Tris-HCl (pH-7.5), 50mM KCl, 1mM DTT and 0.1mM EDTA. 

50mM Tris-HCl, pH7.5 at 25°C, 10mM MgCl₂, 10mM DTT and 1mM ATP. Incubate at 16 °C

Store T4 DNA Ligase at 4°C if entire vial will be used within 2-4 weeks.
Store, frozen at -20°C for longer periods of time.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Avoid multiple freeze-thaw cycles.

1U is the amount of T4 DNA Ligase required to ligate > than 50% DNA fragments in a 20μl ligation reaction system, 6μg of λDNA-Hind III decomposition product reacts at 16°C for 30 minutes.

T4 DNA Ligase can be inhibited by NaCl/KCl at a concentrations >than 200mM or by heating at 65 °C for 10 min or at 70 °C for 5 min.

Cloning of restriction enzyme generated DNA fragments.

Cloning of PCR products.

Joining of double-stranded oligonucleotide linkers or adaptors to DNA.

Site-directed mutagenesis.

Amplified fragment length polymorphism (AFLP).

Ligase-mediated RNA detection.

Nick repair in duplex DNA, RNA or DNA/RNA hybrids.

Self-circularization of linear DNA

SDS