D9380
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:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.53
Recommended Products
grade
for molecular biology
Quality Level
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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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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Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Certificates of Analysis (COA)
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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)
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
Nick translation.
J Meinkoth et al.
Methods in enzymology, 152, 91-94 (1987-01-01)
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