Product Class: Other

OneTaq® Quick-Load® DNA Polymerase
Gray property icon for cloned at NEB recombinant Gray property icon for heat inactivation no 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.

  • Includes green Quick-Load reaction buffer for easy and direct loading of PCR products onto gels
  • 2X higher fidelity than Taq
  • Compatible with standard Taq protocols
Bioz Badge Exists : True
Catalog # Size Concentration
M0509L 500 units 5000 units/ml
M0509X 2500 units 5000 units/ml

Product Information

Description

OneTaq Quick-Load DNA Polymerase is an optimized, ready-to-use blend of Taq and Deep Vent DNA Polymerases 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 convenient quick-load buffer contains two commonly used tracking dyes for direct loading of PCR products on 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 comigrating DNA bands.

 

Figure 1: 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 2: Robust Performance of OneTaq Quick-Load DNA Polymerase

Amplification of a variety of DNA targets (human and viral templates) demonstrates strong performance of the OneTaq Quick-Load DNA Polymerase. Product yield (bars, left axis) and purity (circles, right axis) were calculated via microfluidic analysis from triplicate reactions after 30 cycles of PCR. Standard deviation is indicated by error bars (yield) or shaded bands (purity). GoTaq was cycled according to manufacturer’s recommendations.


Figure 3: 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,
PCR, qPCR & Amplification Technologies,
This product can be used in the following applications:
PCR,
Multiplex PCR,
Specialty PCR,
Routine 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)
    M0509L -20
    Component Name Component # Stored at (°C) Amount Concentration
    OneTaq® Quick-Load® DNA Polymerase M0509LVIAL -20 1 x 0.1 ml 5,000 units/ml
    OneTaq® Standard Reaction Buffer Pack B9022SVIAL -20 2 x 2 ml 5 X
    OneTaq® Quick-Load® Reaction Buffer B0509AVIAL -20 2 x 2 ml 5 X
  • NEB # Stored at (°C)
    M0509X -20
    Component Name Component # Stored at (°C) Amount Concentration
    OneTaq® Quick-Load® DNA Polymerase M0509L -20 5 x 500 units 5,000 units/ml
    OneTaq® Quick-Load® DNA Polymerase M0509LVIAL -20 1 x 0.1 ml 5,000 units/ml
    OneTaq® Standard Reaction Buffer Pack B9022SVIAL -20 2 x 2 ml 5 X
    OneTaq® Quick-Load® Reaction Buffer B0509AVIAL -20 2 x 2 ml 5 X

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.

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

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)

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, 373-374.
  5. Sarkar, G. Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res. 18, 7465.

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Can OneTaq® Quick-Load® DNA Polymerase be used in colony PCR?
  2. What ends will my PCR products have?
  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

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.

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