DNA Ligase T4, 5u/ul

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA. The enzyme repairs single-strand nicks in duplex DNA, RNA or DNA/RNA hybrids, joins DNA fragments with either cohesive or blunt termini (3,4). The T4 DNA Ligase requires ATP as cofactor.\n\nSource:\nE. coli cells carrying a cloned gene 30 from bacteriophage T4.\n\nUnit Definition:\nOne Weiss unit of the enzyme catalyzes the conversion of 1nmol of [32PPi] into Norit-adsorbable form in 20 min. at 37 degrees C (1). One Weiss unit is equivalent to approximately 200 cohesive end ligation units (CEU)*.\nEnzyme activity is assayed in the following mixture: 66mM Tris-HCl, pH 7.6, 6,6mM MgCl2, 0.066mM ATP, 10mM DTT, 3.3uM [32PPi].\n*One CEU is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of lambda DNA in 30 min. at 16 degrees C.\n\nStorage Buffer: \nSupplied in: 20mM Tris-HCl pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 50% glycerol. \n\nSupplied with:\nD3931-01A DNA Ligase T4 Reaction Buffer, 10X: 400mM Tris-HCl, 100mM MgCl2, 100mM DTT, 5mM ATP, pH 7.8\n\nD3931-01B: PEG Solution (50% Polyethylene glycol 4000)

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SPECIFICATIONS

Catalog Number

D3931-01

Size

200U

References

1. Weiss, B., et al., J. Biol. Chem. 243: 4543 (1968) 2. Pheiffer, B.H. & Zimmerman, S.B., Nucleic Acids Res. 11: 7853-7871 (1983). 3. Rossi, R., et al., Nucleic Acids Res. 25: 2106-2113 (1997). 4. Cherepanov, A.V., et al., Biophys. J. 81: 3545-3559 (2001). 5. Nilsson, M., et al., Nucleic Acids Res. 29: 578-581 (2001).

Alternative Names

EC=6.5.1.1; E. coli\nSupplied with Reaction Buffer