Product Class: Other

phi29 DNA Polymerase
NEBU cloned at NEB recombinant 30 65 Heat ralbumin

GMP-grade reagent also available. Learn more.

Product Introduction

Perfect for multiple displacement amplification

  • High fidelity - highly accurate replication
  • Extreme processivity - capable of generating large fragments
  • Isothermal amplification - no need for a thermocycler
Catalog # Size Concentration
M0269S 250.0 units 10000 units/ml
M0269L 1250.0 units 10000 units/ml

Product Information

Description

Highlights

 

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).

Product Source

An E. coli strain that carries the phi29 DNA Polymerase gene from bacteriophage phi29
This product is related to the following categories:
Isothermal Amplification & Strand Displacement Products
This product can be used in the following applications:
Whole Genome Amplification & Multiple Displacement Amplification,
Isothermal Amplification

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 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.

Application Features

  • Replication requiring a high degree of strand displacement and/or processive synthesis
  • High fidelity replication at moderate temperatures

Product Notes

  1. 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

  1. Blanco, L. and Salas, M. (1984). Proc. Natl. Acad. Sci. USA. 81, 5325-5329.
  2. Blanco. L., et al. (1989). J. Biol. Chem.. 264, 8935-8940.
  3. Garmendia, C., et al. (1992). J. Biol. Chem.. 267, 2594-2599.

Protocols, Manuals & Usage

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Can phi29 DNA Polymerase be used in other NEBuffers?
  2. Can phi29 DNA Polymerase be used to blunt DNA?
  3. Can phi29 DNA Polymerase be used to fill in 3' overhangs?
  4. Can phi29 DNA Polymerase be used to remove 5' overhangs?
  5. Can phi29 DNA Polymerase be heat inactivated?
  6. At what rate does phi29 DNA Polymerase add nucleotides to a primed single-stranded template?
  7. Will phi29 DNA Polymerase work in rCutSmart buffer?
  8. Are NEB DNA Polymerases supplied with dNTPs?

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, 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.

Licenses

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.

Trademarks

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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