DNA Polymerase I (Pol I)

DNA Polymerase I (Pol I) catalyzes the incorporation of dNTPs into double-stranded DNA in a 5'→3' direction that is complementary to the DNA or RNA template strand. It possesses 3'→5' exonuclease (proofreading) activity as well as 5'→3' exonuclease activity, making the enzyme useful for DNA end blunting and DNA labeling by nick translation. DNA Polymerase I is supplied in a buffer of 50 mM potassium phosphate (pH 6.5), 1 mM DTT and 50% glycerol.

Applications

DNA labeling by nick translation
DNA end blunting of 5'- and 3'-overhangs
cDNA synthesis from DNA or RNA template

Source

Recombinant E. coli

Storage

–20°C

Unit definition

One unit is defined as the amount of enzyme that catalyzes the incorporation of 10 nmol of total nucleotides into acid-insoluble product in 30 minutes at 37°C and pH 7.4, using poly d(A-T) as the template-primer.

Concentration

3–6 U/µl

Product citations

Friedberg, E. C. The eureka enzyme: the discovery of DNA polymerase. Nat. Rev. Mol. Cell Biol. 7, 143–7 (2006).

Lehman, I. R., Bessman, M. J., Simms, E. S. & Kornberg, A. Enzymatic synthesis of deoxyribonucleic acid. I. Preparation of substrates and partial purification of an enzyme from Escherichia coli. J. Biol. Chem. 233, 163–70 (1958).

Okayama, H. & Berg, P. High-efficiency cloning of full-length cDNA. Mol. Cell. Biol. 2, 161–70 (1982).

Ricchetti, M. & Buc, H. E. coli DNA polymerase I as a reverse transcriptase. EMBO J. 12, 387–96 (1993).

Rigby, P. W., Dieckmann, M., Rhodes, C. & Berg, P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113, 237–51 (1977).

Sambrook, J., Fritsch, E. F. & Maniatis, T. Molecular cloning : a laboratory manual. (Cold Spring Harbor Laboratory, 1989).