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T7 DNA Polymerase (unmodified)

Catalog # Concentration Size List Price Quantity Your Price
M0274L 10000 units/ml 1500 units $461.00
$414.90
M0274S 10000 units/ml 300 units $115.00
$103.50
Catalog # Size List Price Your Price
M0274L 1500 units $461.00
$414.90
M0274S 300 units $115.00
$103.50
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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
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).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  T7 DNA Polymerase (unmodified) M0274SVIAL -20 1 x 0.03 ml 10,000 units/ml
  T7 DNA Polymerase Reaction Buffer B0274AVIAL -20 1 x 1.5 ml 10 X
  Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
  T7 DNA Polymerase (unmodified) M0274LVIAL -20 1 x 0.15 ml 10,000 units/ml
  T7 DNA Polymerase Reaction Buffer B0274AVIAL -20 1 x 1.5 ml 10 X
  Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
Application Features
  • Second strand synthesis in site-directed mutagenesis protocols (8).

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

Companion Products
Materials Sold Separately
Notes
  • The high polymerization rate of the enzyme makes long incubations unnecessary.
  • T7 DNA Polymerase is not suitable for DNA sequencing.
References
  • Hori, K. et al. (1979). J. Biol. Chem.. 254, 11598-11604.
  • Engler, M.J. et al. (1983). J. Biol. Chem.. 258, 11165-11173.
  • Nordstrom, B. et al. (1981). J. Biol. Chem.. 256, 3112-3117.
  • Studier, F.W. et al. (1990). Goeddel, D.V.(Ed.), In Methods Enzymol.. 185, 60-89. San Diego: Academic Press.
  • Grippo, P. and Richardson, C.C. (1971). J. Biol. Chem. 246, 6867-6873.
  • Modrich, P. and Richardson, C.C. (1975). J. Biol. Chem. 250, 5515-5522.
  • Adler, S. and Modrich, P. (1979). J. Biol. Chem. 254, 11605-11614.
  • Bebenek, K. and Kunkel, T.A. (1989). Nucl. Acids Res.. 17, 5408.
  • Mattila, P., et al. (1991). Nucleic Acids Res.. 19, 4967-4973.
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]
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 Data Sheets
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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.SEQUENASE™ is a trademark of GE Healthcare

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