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phi29 DNA Polymerase

Catalog # Concentration Size List Price Quantity Your Price
M0269L 10000 units/ml 1250 units $364.00
$327.60
M0269S 10000 units/ml 250 units $91.00
$81.90
Catalog # Size List Price Your Price
M0269L 1250 units $364.00
$327.60
M0269S 250 units $91.00
$81.90
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Perfect for multiple displacement amplification

  • High fidelity - highly accurate replication
  • Extreme processivity - capable of generating large fragments
  • Isothermal amplification - no need for a thermocycler
phi29 DNA Polymerase is the replicative polymerase from the Bacillus subtilis phage phi29 (Φ29) (1). This polymerase has exceptional strand displacement and processive synthesis properties (2). The polymerase has an inherent 3´→5' proofreading exonuclease activity (3).
Highlights

 

Product Source
An E. coli strain that carries the phi29 DNA Polymerase gene from bacteriophage phi29
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  phi29 DNA Polymerase M0269SVIAL -20 1 x 0.025 ml 10,000 units/ml
  phi29 DNA Polymerase Reaction Buffer B0269SVIAL -20 1 x 1.5 ml 10 X
  Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
  phi29 DNA Polymerase M0269LVIAL -20 1 x 0.125 ml 10,000 units/ml
  phi29 DNA Polymerase Reaction Buffer B0269SVIAL -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
  • Replication requiring a high degree of strand displacement and/or processive synthesis
  • High fidelity replication at moderate temperatures

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 0.5 pmol of dNTP into acid insoluble material in 10 minutes at 30°C.

Reaction Conditions

1X phi29 DNA Polymerase Reaction Buffer
Supplement with Recombinant Albumin, Molecular Biology Grade
Incubate at 30°C

1X phi29 DNA Polymerase Reaction Buffer
50 mM Tris-HCl
10 mM MgCl2
10 mM (NH4)2SO4
4 mM DTT
(pH 7.5 @ 25°C)

Storage Buffer

10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.5% Tween® 20
0.5% IGEPAL® CA-630
pH 7.4 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Theoretical: 67000 daltons

5' - 3' Exonuclease

No

3' - 5' Exonuclease

Yes

Strand Displacement

++++

Unit Assay Conditions

1X phi29 DNA Polymerase Reaction Buffer, 0.1 mg/ml Recombinant Albumin, 0.01 mg/ml HindIII-digested λ DNA, 0.2 µM dTTP including [3H]-dTTP, 0.2 mM dGTP, 0.2 mM dATP and 0.2 mM dCTP.

Companion Products
Materials Sold Separately
Notes
  • The presence of active reducing reagent in the reaction buffer is critical for this enzyme. While the reaction buffer supplied with the enzyme contains DTT, older buffer stocks or stocks that have been repeatedly frozen and thawed should be supplemented with 4 mM DTT to obtain maximal activity.
References
  • Blanco, L. and Salas, M. (1984). Proc. Natl. Acad. Sci. USA. 81, 5325-5329.
  • Blanco. L., et al. (1989). J. Biol. Chem.. 264, 8935-8940.
  • Garmendia, C., et al. (1992). J. Biol. Chem.. 267, 2594-2599.
FAQs
Tech Tips
  • DTT in the reaction buffer can oxidize quickly with >-20°C storage temperatures, extended storage, or multiple freeze/thaws, which can reduce amplification efficiency of phi29 DNA Polymerase.  Older buffer stocks or stocks that have been repeatedly frozen and thawed should be supplemented with 4 mM DTT to obtain maximal activity.
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
Legal And Disclaimer

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.Use of this material in DNA sequencing and/or DNA amplification may require a license from GE Healthcare. Any persons interested in using this product in such applications are advised to contact GE Healthcare, 800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855–1327, Attention: Patent Department.THERMOPOL® is a registered trademark of New England Biolabs, Inc.

IGEPAL® is a registered trademark of Rhodia Operations.
TWEEN® is a registered trademark of Uniqema Americas, LLC.

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