DNA Polymerase I (E. coli)Product information
DNA Polymerase I
|500 units ( 10,000 units/ml )||
DNA Polymerase I
|2.500 units ( 10,000 units/ml )||
DNA Polymerase I (E. coli) is a DNA-dependent DNA polymerase with inherent 3′– 5´ and 5′– 3′ exonuclease activities
- Nick translation of DNA
- Second strand cDNA synthesis
DescriptionDNA Polymerase I (E coli) is a DNA-dependent DNA polymerase with inherent 3´→ 5´ and 5´→ 3´ exonuclease activities (1). The 5´→ 3´ exonuclease activity removes nucleotides ahead of the growing DNA chain, allowing nick-translation.
Product SourceAn E. coli strain that carries an overexpressed copy of the polA gene.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|NEBuffer™ 2||-20||10 X|
Advantages and Features
- Nick translation of DNA to obtain probes with a high specific activity (2)
- Second strand synthesis of cDNA (3,4)
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.
1X NEBuffer™ 2
Incubate at 37°C
1X NEBuffer™ 2
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)
25 mM Tris-HCl
1 mM DTT
0.1 mM EDTA
pH 7.4 @ 25°C
Heat Inactivation75°C for 20 min
Molecular WeightTheoretical: 103000 daltons
5' - 3' ExonucleaseYes
3' - 5' ExonucleaseYes
Unit Assay Conditions1X NEBuffer 2, 33 μM dNTPs including [3H]-dTTP and 70 μg/ml denatured herring sperm DNA.
Error Rate< 9x10-6bases
- DNase I is not included with this enzyme and must be added for nick translation reactions.
- DNA Polymerase I (E.coli) is active in all four NEBuffers when supplemented with dNTPs (not included).
- Lehman, I.R. (1981). In P.D. Boyer(Ed.), The Enzymes. 14A, 16-38. San Diego: Academic Press.
- Meinkoth, J. and Wahl, G.M (1987). S.L. Berger and A.R. Kimmel(Ed.), Methods Enzymol.. 152, 91-94. San Diego: Academic Press.
- Gubler, U. and Hoffmann, B.J. (1983). Gene. 25, 263-269.
- D'Alessio, J.M. and Gerard, G.F. (1988). Nucl. Acids Res.. 16, 1999-2014.
- Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984). J. Biol. Chem. . 259, 1539-1545.
Faqs & Tech Tips
- Can DNA Polymerase I (E. coli) be used in other NEBuffers, including CutSmart?
- Can DNA Polymerase I (E. coli) be used to blunt DNA?
- Can DNA Polymerase I (E. coli) be used to fill in 3' overhangs?
- Why are there artifacts in my blots using the nick translated probe?
- Are NEB DNA Polymerases supplied with dNTPs?
- Can DNA Polymerase I (E. coli) be used to remove 5' overhangs?
- Can DNA Polymerase I (E. coli) be heat inactivated?
- Can DNA Polymerase I (E. coli) be used in nick translation protocols?
- Is nick translation the best way to make a labeled probe?
Protocols & Manuals
Other Tools & Resources
Quality & Safety
Quality Assurance StatementQuality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
SpecificationsThe Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate Of AnalysisThe Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety DataSheetsThe following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
DNA Polymerase I (E. coli)
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.