Description
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.
References
Challberg, M.D. and Englund, P.T. (1980) Meth. Enzymol. 65 , 39–43.
Burd, J.F. and Wells, R.D. (1974) J. Biol. Chem. 249 , 7094–801.
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.
