T4 DNA Polymerase

T4 DNA Polymerase catalyzes the 5´→3´ synthesis of DNA from a primed single-stranded DNA template. Although possessing a potent 3´→5´ proofreading exonuclease, T4 DNA Polymerase contains no 5´→3´ exonuclease activity. T4 DNA Polymerase can be used to fill in 5´ protruding ends with labeled or unlabeled dNTPs or for the generation of blunt ends from DNA molecules with 3´ overhangs or in vitro mutagenesis. The enzyme is provided with 10X Reaction Buffer: 250mM Tris-acetate (pH 7.7), 1M potassium acetate, 100mM magnesium acetate and 10mM DTT.

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References

1. Challberg, M.D. and Englund, P.T. (1980) Meth. Enzymol. 65, 39–43.
2. Burd, J.F. and Wells, R.D. (1974) J. Biol. Chem. 249, 7094–801.
3. Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984) J. Biol. Chem. 259, 1539–45.

Storage Buffer: 200mM potassium phosphate (pH 6.5 at 25°C), 2mM DTT and 50% glycerol.

Source: Recombinant E. coli strain.

QC Tests: Activity, SDS-PAGE/purity, endonuclease.

Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 55nmol of dTTP into acid-precipitable material in 30 minutes at 39°C using poly(dA):oligo(dT) as a substrate.