T4 DNA Ligase

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between neighboring 5’-phosphate and 3’-hydroxyl termini in duplex DNA or RNA. It can be used to join DNA fragments with both sticky ends and blunt ends, and to repair nicks in double-stranded DNA .

Applications

• Cloning of restriction enzyme generated DNA fragments and PCR products
• Next-generation library preparation
• Joining linkers and adapters to cohesive or blunt-ended DNA
• Nick repair in duplex DNA, RNA or DNA/RNA hybrids
• Self-circularization of linear DNA

Product Source

• E. coli strain expressing a recombinant clone

Includes

• T4 DNA Ligase
• 10X T4 DNA Ligase Buffer

10X T4 DNA Ligase reaction buffer

500 mM Tris-HCl, 100 mM MgCl₂, 10 mM ATP, 100 mM DTT, pH 7.5 @ 25ºC

Storage Buffer

50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC

Storage Temperature

-20ºC

Heat Inactivation

65ºC for 10 minutes

Unit Definition

One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA in a total reaction of 20 μl in 30 minutes at 16°C in 1X T4 DNA Ligase reaction buffer.

Quality Control Assays

T4 DNA Ligase is free from detectable nuclease contamination