Product Class: Other

OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer
Gray property icon for cloned at NEB recombinant Gray property icon for heat inactivation no Gray property icon for Hot Start Gray property icon for PCR Gray property icon for annealing temperature -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 nonspecific amplification and does not require a separate activation step
  • Obtain high yields across a wide range of AT / GC content
  • Offered with inert tracking dyes for easy and direct loading of PCR products onto gels
Bioz Badge Exists : True
Catalog # Size Concentration
M0488S 100 reactions (50.0 µl) 2 X
M0488L 500 reactions (50.0 µl) 2 X

Product Information

Description

OneTaq Hot Start Quick-Load 2X Master Mix with Standard 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 routine PCR applications from a variety of 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 Quick-Load master mix formulation contains dNTPs, MgCl2, 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 ~3-5 kb and Tartrazine migrates at ~50 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.


Figure 1: Migration of tracking dyes contained in Quick-Load® PCR reagents during electrophoresis

Green visible tracking dye with Quick-Load® separates to blue ~ 3-5 kb and yellow ~ 50 bp

Quick-Load PCR reagents enable direct loading of PCR products onto agarose DNA gels for analysis.  Quick-Load amplification products deliver the same performance as their non-Quick-Load equivalents, while incorporating two PCR-inert tracking dyes for enhanced visualization. During gel electrophoresis, the blue Xylene Cyanol FF dye co-migrates with DNA fragments of approximately 3-5 kb in a 1% agarose gel, while the yellow Tartrazine dye migrates at approximately 50 bp for clear monitoring of both large and small DNA fragments. Additionally, the combination of the blue and yellow dyes results in a green reagent that enables visual tracking of reaction set up..


[Icon] Use our Tm Calculator to determine your primers' optimal annealing temperature.

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,
Taq DNA Polymerase,
Master Mixes,
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
  • NEB # Stored at (°C)
    M0488S -20
    Component Name Component # Stored at (°C) Amount Concentration
    OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer M0488SVIAL -20 2 x 1.25 ml 2 X
  • NEB # Stored at (°C)
    M0488L -20
    Component Name Component # Stored at (°C) Amount Concentration
    OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer M0488SVIAL -20 10 x 1.25 ml 2 X

Properties & Usage

1X Master Mix Composition

20 mM Tris-HCl
1X Tartrazine
25 units/ml OneTaq Hot Start DNA Polymerase
22 mM KCl
22 mM NH4Cl
1.8 mM MgCl2
5% Glycerol
0.06% IGEPAL® CA-630
0.05% Tween® 20
0.2 mM dNTPs
1X Xylene Cyanol
pH 8.9 @ 25°C

Heat Inactivation

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
  • AT-rich 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 Standard Buffer are available separately.
  2. OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer is stable for fifteen freeze-thaw cycles when stored at -20°C
  3. OneTaq Hot Start Quick-Load 2X Master Mix with Standard 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.
  4. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques. 15, 372-374.
  5. Sarkar, G., Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res.. 18, 7465.

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

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 Standard 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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