Product Class: Other

OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer
cloned at NEB recombinant No hotstart pcr 5

The ONE polymerase for your endpoint PCR needs

Product Introduction

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 nonspecific amplification and does not require a separate activation step

• Obtain high yields with GC rich templates (>65%)

• Offered with inert tracking dyes for easy and direct loading of PCR products onto gels

Catalog # Size Concentration
M0489S 100.0 reactions (50.0 µl) 2 X
M0489L 500.0 reactions (50.0 µl) 2 X

Product Information

Description

Highlights


OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer is an optimized, ready-to-use blend of Taq and Deep Vent® DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to PCR applications from GC-rich templates, including pure DNA solutions, bacterial colonies, and cDNA products. The 3´→5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1). The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient master mix formulation contains dNTPs, MgSO4, buffer components and stabilizers as well as two commonly used tracking dyes for DNA gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~4 kb and Tartrazine migrates at ~10 bp. Both dyes are present in concentrations that do not mask co-migrating DNA bands.

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,
Master Mixes 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
  • M0489S     -20    
      OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer M0489SVIAL -20 2 x 1.25 ml 2 X
      OneTaq® High GC Enhancer B9026AVIAL -20 1 x 2 ml Not Applicable
  • M0489L     -20    
      OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer M0489SVIAL -20 10 x 1.25 ml 2 X
      OneTaq® High GC Enhancer B9026AVIAL -20 3 x 2 ml Not Applicable

Properties & Usage

1X Master Mix Composition

80 mM Tris-SO4
1X Tartrazine
25 units/ml OneTaq Hot Start DNA Polymerase
20 mM (NH4)2SO4
0.2 mM dNTPs
2 mM MgSO4
5% Glycerol
5% DMSO
0.06% IGEPAL® CA-630
0.05% Tween® 20
1X Xylene Cyanol
pH 9.2 @ 25°C

Heat Inactivation

No

Unit Assay Conditions

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

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 Hot Start Quick-Load Master Mix with GC Buffer, DNA template and primers in a total reaction volume of 50 µl.

Application Features

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

Product Notes

  1. Product specifications for individual components in the OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer are available separately.
  2. OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer is stable for fifteen freeze-thaw cycles when stored at -20°C.
  3. OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer is also stable for one month at 4°C, so for frequent use, an aliquot may be kept at 4°C.

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. Protocol for OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer
  2. Guidelines for PCR Optimization with OneTaq® and OneTaq® Hot Start DNA Polymerases

Usage & Guidelines

Application Notes

Tools & Resources

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?

Tech Tips

A master mix with a gel loading dye is the ultimate time-saver! It allows for easy reaction set up and with the gel loading dye already in the mix, the results of your PCR are available even faster.

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 Quick-Load® 2X Master Mix with GC Buffer 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. 

Trademarks

NEW ENGLAND BIOLABS®, ONETAQ®, QUICK-LOAD® 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.

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