DNA Polymerase I (E. coli)

Catalog # Concentration Size List Price Quantity Your Price
M0209L 10000 units/ml 2500 units $454.00
$408.60
M0209S 10000 units/ml 500 units $113.00
$101.70
Catalog # Size List Price Your Price
M0209L 2500 units $454.00
$408.60
M0209S 500 units $113.00
$101.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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
DNA 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 Source
An E. coli strain that carries an overexpressed copy of the polA gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  DNA Polymerase I (E. coli) M0209SVIAL -20 1 x 0.05 ml 10,000 units/ml
  NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
  DNA Polymerase I (E. coli) M0209LVIAL -20 1 x 0.25 ml 10,000 units/ml
  NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
Application Features
  • Nick translation of DNA to obtain probes with a high specific activity (2)
  • Second strand synthesis of cDNA (3,4)

Properties & Usage

Unit Definition

One 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.

Reaction Conditions

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)

Storage Buffer

25 mM Tris-HCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

75°C for 20 minutes

Molecular Weight

Theoretical: 103000 daltons

5' - 3' Exonuclease

Yes

3' - 5' Exonuclease

Yes

Strand Displacement

No

Unit Assay Conditions

1X NEBuffer 2, 33 μM dNTPs including [3H]-dTTP and 70 μg/ml denatured herring sperm DNA.

Error Rate

<>-6bases

Materials Sold Separately
Notes
  • 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).
References
  • 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.
Quality Control Assay
Quality 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.
Specifications
The 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]
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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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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