Product Class: Other

DNA Polymerase I (E. coli)
NEB2 cloned at NEB recombinant 37 75 Heat

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
M0209S 500 units 10000 units/ml
M0209L 2500 units 10000 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.
This product is related to the following categories:
DNA Labeling,
DNA Manipulation,
cDNA Synthesis & Reverse Transcriptases,
This product can be used in the following applications:
Polymerases for DNA Manipulation,
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

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

< 9x10-6bases

Application Features

  • Nick translation of DNA to obtain probes with a high specific activity (2)
  • Second strand synthesis of cDNA (3,4)

Product 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.

Protocols, Manuals & Usage

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Which DNA polymerase can I use to blunt DNA?
  2. Can I use a DNA polymerase to fill in 3' overhangs or remove 5' overhangs?
  3. How can I heat inactivate this DNA polymerase?
  4. Can this DNA polymerase be used in labeling and partial fill reactions?
  5. Can this DNA polymerase be used in other NEBuffers for filling in or blunting overhangs?

Quality, Safety & Legal

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 controls 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.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].

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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