Description:
T4 DNA Polymerase catalyzes the extension of a primed DNA template in the 5' -> 3' direction. This enzyme exhibits a powerful 3' -> 5' exonuclease activity, while lacking any inherent 5' -> 3' exonuclease or strand displacement functions.
Specific Activity: 5,555U/mg
Applications:
-
3'-overhang removal to form blunt ends
-
5'-overhang fill-in to form blunt ends
-
Single strand deletion for sub-cloning
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Second strand synthesis in site-directed mutagenesis
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Probe labeling using replacement synthesis
Source:
Purified from a strain of E. coli that expresses the recombinant T4 DNA Polymerase gene.
Supplied in:
100 mM KPO4
1.0 mM DTT
0.1 mM EDTA
50% Glycerol
pH 6.5 @ 25°C
Supplied With: 10x Blue Buffer
10x Blue Buffer:
500 mM NaCl
100 mM Tris-HCl
100 mM MgCl2
10 mM DTT
pH 7.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid-precipitable material in 30 minutes at 37°C.
Recommended Storage Condition: -20ºC
Reference:
1. Tabor, S. and Struhl, K. (1989) In DNA-Dependent DNA Polymerases. F. M. Ausebel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl (Eds.), Current Protocols in Molecular Biology, pp. 3.5.10-3.5.12. 2. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, (2nd Ed.), 5.44-5.47.
2. Dale, R., McClure, B. and Houchins, J. (1985) Plasmid, 13, 31-40.
3. Kunkel, T.A., Roberts, J.D. and Zakour, R.A. (1987) R. Wu and L. Grossman (Eds.), Methods Enzymol., 154, pp. 367-382. San Diego: Academic Press.
4. Panet, A., van de Sande, J.H., Loewen, P.C. and Khorana, H.G. (1973) Biochemistry, 12, 5045-5050.
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