Description
Biochem/physiol Actions DNA polymerase I （holoenzyme） has 5′→3′ and 3′→5′ exonuclease activities in addition to its synthetic activity. This bifunctional activity enables the enzyme to use nicks or gaps in double stranded DNA as starting points for DNA synthesis. The 5′→3′ exonuclease activity degrades the DNA strand complementary to the template strand beginning at the nick. DNA synthesis begins at the 3′-end of the nick and produces a new strand of DNA complementary to the template. The net result is the movement of the polymerase along the template strand （nick translation） until all the DNA complementary to the template （from the site of the original nick to the 5′-end of the template strand） is replaced. The enzyme is used with radioactive or biotinylated nucleotides to prepare labeled DNA of high specific activity.
DNA polymerase I has also been reported to catalyze de novo DNA synthesis.
Features and Benefits • Highly specific DNA probes by nick translation.
• In vitro synthesis of complementary cDNA strand.
• In vitro synthesis of DNA.
• Produce blunt ends from 5′ and 3′ overhangs
Unit Definition One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min at 37 °C.
Physical form Solution in 50% glycerol containing 100 mM potassium phosphate buffer, pH 6.5, and 1 mM dithiothreitol
Properties
form buffered aqueous glycerol solution
concentration 5,000-15,000 units/mL
foreign activity Endonuclease, none detected
shipped in wet ice
storage temp. −20°C
Safety
Personal Protective Equipment Eyeshields, Gloves, half-mask respirator （EU）, half-mask respirator （US）, multi-purpose combination respirator cartridge （US）
Safety Statements 24/25
WGK Germany 3
References
reference Lehman, I.R.,, Boyer, P.D., ed. The Enzymes San Diego, CA , （1981） 14A, 16-38
Meinkoth, J., and Wahl, G.M., Nick translation. Meth. Enzymol. 152, 91-94, （1987） Abstract
Maniatis, T., et al., Nucleotide sequence of the rightward operator of phage lambda. Proc. Natl. Acad. Sci. U. S. A. 72, 1184-1188, （1975） Abstract
Gubler, U., et al., A simple and very efficient method for generating cDNA libraries. Gene 25, 263-269, （1983） Abstract
D'Alessio, J.M. and Gerard, G.F., Second-strand cDNA synthesis with E. coli DNA polymerase I and RNase H: the fate of information at the mRNA 5' terminus and the effect of E. coli DNA ligase. Nucleic Acids Res. 16, 1999-2014, （1988） Abstract
Harwood, S.J., et al., Micrococcus luteus deoxyribonucleic acid polymeraseStudies of the enzymic reaction and properties of the deoxyribonucleic acid product. J. Biol. Chem. 245, 5614-5624, （1970） Abstract
Okayama, H., and Berg, P., High-efficiency cloning of full-length cDNA. Mol. Cell. Biol. 2, 161-170, （1982） Abstract
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