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T4 DNA Polymerase

Catalog # Concentration Size List Price Quantity Your Price
M0203L 3000 units/ml 750 units $416.00
$374.40
M0203S 3000 units/ml 150 units $104.00
$93.60
Catalog # Size List Price Your Price
M0203L 750 units $416.00
$374.40
M0203S 150 units $104.00
$93.60
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

T4 DNA Polymerase catalyzes the synthesis of DNA in the 5´→ 3´ direction and requires the presence of template and primer. 

  • Gap filling (no strand displacement activity)
  • Removal of 3’ overhangs or fill-in of 5’ overhangs to form blunt ends
  • Lacks 5’ —> 3’ exonuclease activity
  • Probe labeling using replacement synthesis
  • Singe-strand deletion subcloning
Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    DNA Blunting Tutorial
  • Monarch_MiniprepProtocol_VideoThumb
    Traditional Cloning Workflow
T4 DNA Polymerase catalyzes the synthesis of DNA in the 5´→ 3´ direction and requires the presence of template and primer. This enzyme has a 3´→ 5´ exonuclease activity which is much more active than that found in DNA Polymerase I (E. coli).  Unlike E. coli DNA Polymerase I, T4 DNA Polymerase does not have a 5´→ 3´ exonuclease function.
Product Source
Purified from a strain of E. coli that carries the T4 DNA Polymerase gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  T4 DNA Polymerase M0203SVIAL -20 1 x 0.05 ml 3,000 units/ml
  NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X
  T4 DNA Polymerase M0203LVIAL -20 1 x 0.25 ml 3,000 units/ml
  NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X
Application Features
  • 3´ overhang removal to form blunt ends (1,2).
  • 5´ overhang fill-in to form blunt ends (1,2).
  • Single strand deletion subcloning (3).
  • Second strand synthesis in site-directed mutagenesis (4).
  • Probe labeling using replacement synthesis (1,2).

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 (5).

Reaction Conditions

1X NEBuffer™ r2.1

1X NEBuffer™ r2.1
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBufferâ„¢ 1: 60%
NEBufferâ„¢ 2: 100%
NEBufferâ„¢ 3: 100%
NEBufferâ„¢ 4: 100%

Storage Buffer

100 mM KPO4
1 mM DTT
50% Glycerol
pH 6.5 @ 25°C

Heat Inactivation

75°C for 20 minutes

Molecular Weight

Theoretical: 104000 daltons

5' - 3' Exonuclease

No

3' - 5' Exonuclease

Yes

Strand Displacement

No

Unit Assay Conditions

1X NEBuffer 2.1, 33 µM dNTPs including [3H]-dTTP, 70 µg/ml denatured herring sperm DNA.

Error Rate

~ 1x10-6bases

Companion Products
Materials Sold Separately
Notes
  • For fill-in reactions only: T4 DNA Polymerase can be used in NEBuffers 1.1, 2.1, 3.1 and CutSmart® Buffer as well as NEBuffers 1-4 and T4 DNA Ligase Reaction Buffer.
  • For blunting reactions requiring removal of overhangs: T4 DNA Polymerase can be used in NEBuffers 1.1, 2.1, and CutSmart Buffer as well as NEBuffers 1, 2, and 4 and T4 DNA Ligase Reaction Buffer. NEBuffers 3.1 and 3 are not recommended when overhang removal is required.
  • Optimal activity is observed in NEBuffer 2.1.
  • Supplement with dNTPs.*
  • BSA supplementation is recommended when using a buffer that does not already contain BSA.
  • Incubate at temperature suggested for specific protocol.

    * Refer to specific protocol to determine recommended dNTP concentrations.
    Refer
  • Elevated temperatures, excessive amounts of enzyme, failure to supplement with dNTPs or long reaction times may result in recessed ends due to the 3´→ 5´ exonuclease activity of the enzyme.
References
  • Tabor, S. and Struhl, K. (1989). DNA-Dependent DNA Polymerases. In F. M. Ausebel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl(Ed.), Current Protocols in Molecular Biology. 3.5.10-3.5.12. New York: John Wiley & Sons, Inc.
  • Sambrook, J. et al. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.44-5.47. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
  • Dale, R. et al. (1985). Plasmid. 13, 31-40.
  • Kunkel, T.A. et al. (1987). Methods Enzymol.. 154, 367-382.
  • Panet, A. et al. (1973). Biochemistry. 12, 5045-5050.
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.THERMOPOL® is a registered trademark of New England Biolabs, Inc.

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