T4 DNA Ligase
T4 DNA Ligase
T4 DNA Ligase is a ligation enzyme that can be used to join DNA fragments by catalyzing the formation of phosphodiester bonds between juxtaposed 5' phosphate and 3' hydroxyl termini in double-stranded DNA using ATP as a coenzyme. Both blunt and cohesive end DNA ligation, as well as single-stranded nick repair of DNA, RNA and DNA/RNA, are possible using the T4 DNA ligase.
Applications
- Blunt and cohesive end DNA ligation
- Blunt end linker addition
- Single-strand DNA, RNA and DNA/RNA repair
- DNA fragment insertion into vectors
Source
Recombinant E. coli
Storage
–20°C
Unit definition
One ligation unit is defined as the amount of enzyme that ligates >90% of 6 µg of Hind III-restricted lambda DNA in a 20 µl reaction volume in 30 minutes at 16°C. One unit of the enzyme corresponds to 0.008 Weiss units by the ATP-PPi exchange reaction in 66 mM Tris-HCl (pH 7.6), 6.6 mM MgCl2, 10 mM DTT, 66 µM ATP and 3.3 µM [32P]Na4P2O7.
Concentration
350 U/µl (2.8 Weiss units/µl)
Product citations
Weiss, B., Jacquemin-Sablon, A., Live, T. R., Fareed, G. C. & Richardson, C. C. Enzymatic breakage and joining of deoxyribonucleic acid. VI. Further purification and properties of polynucleotide ligase from Escherichia coli infected with bacteriophage T4. J. Biol. Chem. 243, 4543–55 (1968).
Weiss, B., Thompson, A. & Richardson, C. C. Enzymatic breakage and joining of deoxyribonucleic acid. VII. Properties of the enzyme-adenylate intermediate in the polynucleotide ligase reaction. J. Biol. Chem. 243, 4556–63 (1968).
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