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.
- DNA labeling by nick translation
- DNA end blunting of 5'- and 3'-overhangs
- cDNA synthesis from DNA or RNA template
Recombinant E. coli
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.
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).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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