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D9380

Sigma-Aldrich

DNA Polymerase I from Escherichia coli lysogenic for NM 964

buffered aqueous glycerol solution

Synonym(s):

Kornberg Polymerase

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About This Item

CAS Number:
9012-90-2
Enzyme Commission number:
2.7.7.7 (BRENDA, IUBMB)
MDL number:
MFCD00130924
UNSPSC Code:
12352204
NACRES:
NA.53

grade

for molecular biology

Quality Level

200

form

buffered aqueous glycerol solution

mol wt

109 kDa

concentration

5,000-15,000 units/mL

UniProt accession no.

P00582

foreign activity

Endonuclease, none detected

shipped in

wet ice

storage temp.

−20°C

Gene Information

Escherichia coli K12 ... polA(948356)

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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.

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LIG1

DNA Ligation Kit

pictograms

Health hazard
GHS08

signalword

Danger

hcodes

H334

Hazard Classifications

Resp. Sens. 1

Storage Class

10 - Combustible liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)

Certificates of Analysis (COA)

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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)
Lehman, I.R., et al.
The Enzymes, 14A, 16-38 (1981)
H Okayama et al.
Molecular and cellular biology, 2(2), 161-170 (1982-02-01)
A widely recognized difficulty of presently used methods for cDNA cloning is obtaining cDNA segments that contain the entire nucleotide sequence of the corresponding mRNA. The cloning procedure described here mitigates this shortcoming. Of the 10(5) plasmid-cDNA recombinants obtained per
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
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)
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