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D9380

Sigma-Aldrich

DNA Polymerase I from Escherichia coli lysogenic for NM 964

buffered aqueous glycerol solution

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Synonym(s):
Kornberg Polymerase
CAS Number:
Enzyme Commission number:
MDL number:
NACRES:
NA.53

grade

for molecular biology

form

buffered aqueous glycerol solution

mol wt

109 kDa

concentration

5,000-15,000 units/mL

UniProt accession no.

foreign activity

Endonuclease, none detected

shipped in

wet ice

storage temp.

−20°C

Gene Information

Escherichia coli K12 ... polA(948356)

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This Item
D8276D1806D4545
form

buffered aqueous glycerol solution

form

buffered aqueous glycerol solution

form

liquid

form

liquid

mol wt

109 kDa

mol wt

103 kDa

mol wt

94 kDa

mol wt

-

concentration

5,000-15,000 units/mL

concentration

~3,000 units/mL

concentration

5 units/μL

concentration

5 units/μL

UniProt accession no.

P00582

UniProt accession no.

P00582

UniProt accession no.

-

UniProt accession no.

-

foreign activity

Endonuclease, none detected

foreign activity

Endonuclease, none detected

foreign activity

-

foreign activity

-

General description

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 the DNA complementary to the template (from the site of the original nick to the 5′-end of the template strand) is replaced.

Application

DNA Polymerase I from Escherichia coli has been used to study the effects of the anti-tumor drug cis-diaminedichloroplatinum (II) on the enzyme activity.
Suitable for:
  • 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

Components

DNase Polymerase I is supplied in a solution of 50% glycerol containing 100 mM potassium phosphate buffer (pH 6.5), and 1 mM dithiothreitol.

Unit Definition

One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min at 37 °C.

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Micrococcus luteus deoxyribonucleic acid polymerase. Studies of the enzymic reaction and properties of the deoxyribonucleic acid product.
S J Harwood et al.
The Journal of biological chemistry, 245(21), 5614-5624 (1970-11-10)
U Gubler et al.
Gene, 25(2-3), 263-269 (1983-11-01)
A simple method for generating cDNA libraries from submicrogram quantities of mRNA is described. It combines classical first-strand synthesis with the novel RNase H-DNA polymerase I-mediated second-strand synthesis [Okayama, H., and Berg, P., Mol. Cell. Biol. 2 (1982) 161-170]. Neither
J M D'Alessio et al.
Nucleic acids research, 16(5), 1999-2014 (1988-03-25)
A simple method for generating cDNA libraries has been described (1) in which RNase H-DNA polymerase I-mediated second-strand cDNA synthesis primes from an RNA oligonucleotide derived from the 5' (capped) end of mRNA. The size of this oligonucleotide and the
Lehman, I.R., et al.
The Enzymes, 14A, 16-38 (1981)
Inhibition of Escherichia coli DNA polymerase-I by the anti-cancer drug cis-diaminedichloroplatinum(II): what roles do polymerases play in cis-platin-induced cytotoxicity?
Rebecca K
Febs Letters (1999)

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