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