DNA Polymerase I catalyzes the template-directed polymerization of nucleotides into duplex DNA in a 5´→3´ direction. DNA Polymerase I possesses a 3´→5´ exonuclease activity or "proofreading" function, which lowers the error rate during DNA replication, and also contains a 5´→3´ exonuclease activity, which enables the enzyme to replace nucleotides in the growing strand of DNA by nick translation (1). The enzyme, purified from recombinant E. coli, is capable of catalyzing de novo synthesis of synthetic homopolymers and provides a convenient method for the preparation of a variety of defined DNA substrates (2).
Flexible: DNA Polymerase I may be used in a variety of molecular applications.
May Be Heat-Inactivated: DNA Polymerase I is inactivated by heating at 68°C for 10 minutes.
Provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C), 100mM MgSO4, 1mM DTT.
Labeling of DNA to high specific radioactivity by nick translation (1).
Second-strand cDNA synthesis.
Kelly, R.B. et al. (1970) J. Biol. Chem.245, 39–45.
Harwood, S.J. et al. (1970) J. Biol. Chem.245, 5614–24.