Product Class: Other

T7 DNA Polymerase (unmodified)
NEBU cloned at NEB recombinant 37 75 Heat ralbumin

Product Introduction

T7 DNA Polymerase catalyzes the replication of T7 phage DNA during infection. The high polymerization rate of the enzyme makes long incubations unnecessary.

  • Second strand synthesis in site-directed mutagenesis protocols
  • Gap filling reaction (no strand displacement)
  • T7 DNA Polymerase is not suitable for DNA sequencing
  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
Catalog # Size Concentration
M0274S 300.0 units 10000 units/ml
M0274L 1500.0 units 10000 units/ml

Product Information

Description

T7 DNA Polymerase catalyzes the replication of T7 phage DNA during infection. The protein dimer has two catalytic activities: DNA polymerase activity and strong 3´→ 5´ exonuclease (1,2,3). The high fidelity and rapid extension rate of the enzyme make it particularly useful in copying long stretches of DNA template.

Product Source

T7 DNA Polymerase consists of two subunits: T7 gene 5 protein (84 kilodaltons) and E.coli thioredoxin (12 kilodaltons) (1,4-7). Each protein is cloned and overexpressed in a T7 expression system in E. coli (4).
This product is related to the following categories:
DNA Manipulation Products
This product can be used in the following applications:
Isothermal Amplification,
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 incorporate10 nmoles of dNTP into acid insoluble material in 30 minutes at 37°C.

Reaction Conditions

1X T7 DNA Polymerase Reaction Buffer
Supplement with Recombinant Albumin, Molecular Biology Grade
Incubate at 37°C

1X T7 DNA Polymerase Reaction Buffer
20 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.5 @ 25°C)

Storage Buffer

50 mM KPO4
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7 @ 25°C

Heat Inactivation

75°C for 20 minutes

5' - 3' Exonuclease

No

3' - 5' Exonuclease

Yes

Strand Displacement

No

Unit Assay Conditions

100 mM KCl, 20 mM Tris-HCl (pH 7.6), 6 mM MgCl2 , 0.5 mM DTT, 0.1 mM EDTA, 50 ug/ml Recombinant Albumin, 150 μM dNTPs including [3H]-dTTP and 70 µg/ml denatured herring sperm DNA.

Error Rate

~ 15x10-6bases

Application Features

  • Second strand synthesis in site-directed mutagenesis protocols (8).

Product Notes

  1. The high polymerization rate of the enzyme makes long incubations unnecessary.
  2. T7 DNA Polymerase is not suitable for DNA sequencing.

References

  1. Hori, K. et al. (1979). J. Biol. Chem.. 254, 11598-11604.
  2. Engler, M.J. et al. (1983). J. Biol. Chem.. 258, 11165-11173.
  3. Nordstrom, B. et al. (1981). J. Biol. Chem.. 256, 3112-3117.
  4. Studier, F.W. et al. (1990). Goeddel, D.V.(Ed.), In Methods Enzymol.. 185, 60-89. San Diego: Academic Press.
  5. Grippo, P. and Richardson, C.C. (1971). J. Biol. Chem. 246, 6867-6873.
  6. Modrich, P. and Richardson, C.C. (1975). J. Biol. Chem. 250, 5515-5522.
  7. Adler, S. and Modrich, P. (1979). J. Biol. Chem. 254, 11605-11614.
  8. Bebenek, K. and Kunkel, T.A. (1989). Nucl. Acids Res.. 17, 5408.
  9. Mattila, P., et al. (1991). Nucleic Acids Res.. 19, 4967-4973.

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. What are the main causes of blunting reaction failure?
  6. Can this DNA polymerase be used in other NEBuffers for filling in or blunting overhangs?
  7. Does NEB carry modified T7 Polymerase?

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 busdev@neb.com.

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.

Trademarks

SEQUENASE™ is a trademark of GE Healthcare

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