Product Class: Other

OneTaq® DNA Polymerase
NEBU cloned at NEB recombinant No pcr 5

Product Introduction

The ONE polymerase for your endpoint PCR needs. OneTaq DNA Polymerase is an optimized blend of Taq and Deep Vent® DNA polymerases for use with routine and difficult PCR experiments.

  • 2X higher fidelity than Taq
  • Ideal for routine, AT- or GC-rich templates
  • Hot start and master mix versions available
Catalog # Size Concentration
M0480S 200.0 units 5000 units/ml
M0480L 1000.0 units 5000 units/ml
M0480X 5000.0 units 5000 units/ml

Product Information

Description

The 3´→ 5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1) . The OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields with minimal optimization, regardless of a template’s GC content.

OneTaq DNA Polymerase is supplied with two 5X buffers: (Standard and GC), as well as a High GC Enhancer solution. For most routine and/or AT-rich amplicons (Lambda, etc.) or complex amplicons with up to ~65% GC content, OneTaq Standard Reaction Buffer provides robust amplification. For GC-rich amplicons, the OneTaq GC Reaction Buffer can improve both performance and yield. For particularly high GC or difficult amplicons, the OneTaq High GC Enhancer can be added at a final concentration of 10–20% to reactions containing OneTaq GC Reaction Buffer.

Comparison of OneTaq Products to Other Commercially Available Polymerases

OneTaq_ComparisonChart_0220 
Amplification of a selection of high GC human genomic DNA targets demonstrates OneTaq performance. All polymerases were cycled according to manufacturer’s recommendations, including the use of additives to enhance the amplification of targets with high GC content. Yield (dot size) and purity (color) of reaction product were quantified from triplicate reactions on a Perkin Elmer LabChip®. A large, dark green dot represents the highest yield and purity.


pcr amplification, amplification ,dna amplification

 

Product Source

An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1 and an E. coli strain that carries the Deep Vent® DNA Polymerase gene from Pyrococcus species GB-D.
This product is related to the following categories:
OneTaq® DNA Polymerases Products,
Taq DNA Polymerase Products,
This product can be used in the following applications:
Multiplex PCR,
Specialty PCR,
Routine PCR,
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0480S     -20    
  • M0480L     -20    
  • M0480X     -20    

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 15 nmol of dNTP into acid insoluble material in 30 minutes at 75°C.

Reaction Conditions

1X OneTaq® Standard Reaction Buffer Pack

1X OneTaq® Standard Reaction Buffer Pack
20 mM Tris-HCl
22 mM NH4Cl
22 mM KCl
1.8 mM MgCl2
0.06% IGEPAL® CA-630
0.05% Tween® 20
(pH 8.9 @ 25°C)

Usage Concentration

1.25 units/50μl reaction

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

No

5' - 3' Exonuclease

Yes

3' - 5' Exonuclease

Yes

Strand Displacement

No

Unit Assay Conditions

1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA

Error Rate

< 140x10-6bases

Features

  • Ideal for routine, AT- and GC-rich PCR

Application Features

  • High Sensitivity PCR
  • High Throughput PCR 
  • Routine PCR
  • GC-rich PCR
  • AT-rich PCR
  • Colony PCR 
  • Long PCR (up to ~6 kb genomic)

Product Notes

  1. The OneTaq High GC Enhancer should not be used alone. It should be added only to reactions with the OneTaq GC Reaction Buffer and will typically improve yields when other conditions have failed.
  2. Product specifications for individual components in the OneTaq DNA Polymerase mix are available separately.

References

  1. Barnes, W.M. (1994). Proc. Natl. Acad. Sci. USA. 91, 2216-2220.
  2. Saiki R.K. et al. (1985). Science. 230, 1350-1354.
  3. Powell, L.M. et al. (1987). Cell. 50, 831-840.

Protocols, Manuals & Usage

Protocols

  1. PCR Protocol for OneTaq® DNA Polymerase (M0480)

Usage & Guidelines

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What ends will my PCR products have?
  2. How should I determine the appropriate annealing temperature for my reaction?
  3. What should the final primer concentration be in my PCR?
  4. What are the properties of this polymerase (fidelity, product ends, max amplicon, modified base incorporation, etc.)?
  5. What are the stability and storage requirements for NEB's Taq and OneTaq DNA Polymerases?
  6. Can Taq/OneTaq® DNA Polymerases be used for nick translation?
  7. How can I optimize my PCR when using Taq DNA Polymerase?

Troubleshooting

Tech Tips

Did you know most OneTaq reactions amplify more efficiently and robustly when you use a 68°C extension temperature?

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

Purchase of this product provides the purchaser with a non-exclusive license to use OneTaq® DNA Polymerase for research purposes only.

NEW ENGLAND BIOLABS®, ONETAQ® and THERMOPOL® are registered trademarks of New England Biolabs, Inc.
DEEP VENT® is a trademark of New England Biolabs, Inc.
IGEPAL® is a registered trademark of Rhodia Operations.
TWEEN® is a registered trademark of Uniqema Americas, LLC.