Product Class: Other

DNA Polymerase I (E. coli)

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Product Introduction

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
Catalog # Size Concentration
M0209L 2,500 units 10,000 units/ml
M0209S 500 units 10,000 units/ml

Product Information

Description

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:

Store at (°C) Concentration
NEBuffer™ 2 -20 10 X
Product Categories:
DNA Labeling,
DNA Manipulation,
cDNA Synthesis & Reverse Transcriptases
Applications:
Polymerases for DNA Manipulation,
RT-qPCR, RT-PCR and cDNA Synthesis,
PCR

Advantages and Features

Application 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 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 Temperature

-20°C

Storage Conditions

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

Heat Inactivation

75°C for 20 min

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

< 9x10-6bases

Notes

  1. DNase I is not included with this enzyme and must be added for nick translation reactions.
  2. DNA Polymerase I (E.coli) is active in all four NEBuffers when supplemented with dNTPs (not included).

References

  1. Lehman, I.R. (1981). In P.D. Boyer(Ed.), The Enzymes. 14A, 16-38. San Diego: Academic Press.
  2. Meinkoth, J. and Wahl, G.M (1987). S.L. Berger and A.R. Kimmel(Ed.), Methods Enzymol.. 152, 91-94. San Diego: Academic Press.
  3. Gubler, U. and Hoffmann, B.J. (1983). Gene. 25, 263-269.
  4. D'Alessio, J.M. and Gerard, G.F. (1988). Nucl. Acids Res.. 16, 1999-2014.
  5. Kunkel, T.A., Loeb, L.A. and Goodman, M.F. (1984). J. Biol. Chem. . 259, 1539-1545.

Faqs & Tech Tips

FAQs

  1. Can DNA Polymerase I (E. coli) be used in other NEBuffers, including CutSmart?
  2. Can DNA Polymerase I (E. coli) be used to blunt DNA?
  3. Can DNA Polymerase I (E. coli) be used to fill in 3' overhangs?
  4. Why are there artifacts in my blots using the nick translated probe?
  5. Are NEB DNA Polymerases supplied with dNTPs?
  6. Can DNA Polymerase I (E. coli) be used to remove 5' overhangs?
  7. Can DNA Polymerase I (E. coli) be heat inactivated?
  8. Can DNA Polymerase I (E. coli) be used in nick translation protocols?
  9. Is nick translation the best way to make a labeled probe?

Protocols & Manuals

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.)

Other Tools & Resources

Selection Charts

Interactive Tools

Quality & Safety

Quality Assurance Statement

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]

Certificate Of Analysis

The 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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.