Product Class: Other

OneTaq® Hot Start DNA Polymerase
NEBU cloned at NEB recombinant No hotstart 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.

  • Unique hot start formulation eliminates non-specific amplification and does not require a separate activation step
  • Robust yield with minimal optimization
  • Ideal for routine, AT- or GC-rich templates
  • Room temperature reaction setup
Catalog # Size Concentration
M0481S 200.0 units 5000 units/ml
M0481L 1000.0 units 5000 units/ml
M0481X 5000.0 units 5000 units/ml

Featured Videos

View Video Library
  • Amplify_GC_rich_DNA_TN

    How to Amplify GC-rich DNA

  • PCRTips_VideoThumb

    5 Tips for Setting Up Your PCR

  • TmImportance_VideoThumb

    Why is Tm Important in Primer Design?

  • PCROverview_thumb

    Overview of PCR

Product Information

Description

OneTaq® Hot Start DNA Polymerase is an optimized blend of Taq and Deep Vent® DNA polymerases combined with an aptamer-based inhibitor. The 3´→ 5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1) and the hot start formulation combines convenience with decreased interference from primer dimers and secondary products. The OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields with minimal optimization, regardless of a template’s GC content.

The inhibitor binds reversibly, blocking polymerase activity at temperatures below 45°C. The polymerase is activated during normal cycling conditions, allowing reactions to be set up at room temperature. OneTaq Hot Start DNA Polymerase does not require a separate high temperature incubation step to activate the enzyme and can be used in typical Taq-based cycling protocols.

OneTaq Hot Start 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 (i.e. ≥ 75% GC), 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.


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:
Hot Start PCR,
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
  • M0481S     -20    
      OneTaq® Hot Start DNA Polymerase M0481SVIAL -20 1 x 0.04 ml 5,000 units/ml
      OneTaq® Standard Reaction Buffer Pack B9022SVIAL -20 1 x 2 ml 5 X
      OneTaq® GC Reaction Buffer Pack B9023SVIAL -20 1 x 2 ml 5 X
      OneTaq® High GC Enhancer B9026AVIAL -20 1 x 2 ml Not Applicable
  • M0481L     -20    
      OneTaq® Hot Start DNA Polymerase M0481LVIAL -20 1 x 0.2 ml 5,000 units/ml
      OneTaq® Standard Reaction Buffer Pack B9022SVIAL -20 5 x 2 ml 5 X
      OneTaq® GC Reaction Buffer Pack B9023SVIAL -20 3 x 2 ml 5 X
      OneTaq® High GC Enhancer B9026AVIAL -20 3 x 2 ml Not Applicable
  • M0481X     -20    
      OneTaq® Hot Start DNA Polymerase M0481L -20 5 x 1,000 units 5,000 units/ml
      OneTaq® Hot Start DNA Polymerase M0481LVIAL -20 1 x 0.2 ml 5,000 units/ml
      OneTaq® Standard Reaction Buffer Pack B9022SVIAL -20 5 x 2 ml 5 X
      OneTaq® GC Reaction Buffer Pack B9023SVIAL -20 3 x 2 ml 5 X
      OneTaq® High GC Enhancer B9026AVIAL -20 3 x 2 ml Not Applicable

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. 

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 the OneTaq GC Reaction Buffer and will typically enhance yields when other conditions have failed.
  2. Product specifications for individual components in the OneTaq Hot Start 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, 1530-1554.
  3. Powell, L.M. et al. (1987). Cell. 50, 831-840.

Protocols, Manuals & Usage

Protocols

  1. Protocol for OneTaq Hot Start DNA Polymerase (M0481)

Usage & Guidelines

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

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

Troubleshooting

Tech Tips

Did you know most OneTaq Hot Start 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

Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the OneTaq® Hot Start DNA Polymerase for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com. 

Other Products You May Be Interested In