Product Class: Other

OneTaq® DNA Polymerase

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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
M0480L 1,000 units 5,000 units/ml
M0480S 200 units 5,000 units/ml
M0480X 5,000 units 5,000 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.

pcr amplification, amplification ,dna amplification
pcr amplification, amplification ,dna amplification
Amplification of a selection of sequences with varying GC content from human and C. elegans genomic DNA using OneTaq DNA Polymerase. GC content is indicated above gel. Marker M is the 1 kb DNA Ladder (NEB #N3232).

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 VentR DNA Polymerase gene from Pyrococcus species GB-D.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
OneTaq® Standard Reaction Buffer Pack -20 5 X
OneTaq® GC Reaction Buffer Pack -20 5 X
OneTaq® High GC Enhancer -20 5 X
Product Categories:
OneTaq® DNA Polymerase Products,
Taq DNA Polymerase Products
Applications:
Multiplex PCR,
PCR,
Specialty PCR
,
Routine PCR

Advantages and Features

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)

Properties and 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 Temperature

-20°C

Storage Conditions

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

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.

Faqs & Tech Tips

FAQs

  1. Can I use my regular Taq-based cycling conditions for OneTaq® DNA Polymerase based products?
  2. Which buffer should I use?
  3. When should I add the High GC Enhancer?
  4. Can OneTaq® DNA Polymerase be used in colony PCR?
  5. How long a product can be made by OneTaq® DNA Polymerase?
  6. Can OneTaq® DNA Polymerase be used with uracil-containing primers or bisulfite-treated DNA?
  7. What is the fidelity of OneTaq® DNA Polymerase?
  8. Where can I find help troubleshooting my PCR?
  9. How should I determine an appropriate annealing temperature for my reaction?
  10. Which buffer should I use if I want to control the level of magnesium in the reaction?
  11. How is OneTaq DNA Polymerase different from LongAmp™ Taq DNA Polymerase?
  12. How should I set up an amplification reaction using OneTaq® DNA Polymerase?
  13. What type of DNA ends result from a primer extension reaction or a PCR using OneTaq® DNA Polymerase?

Tech Tips

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

Protocols & Manuals

Protocols

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

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.)

Other Tools & Resources

Selection Charts

Usage Guildelines & Tips

Troubleshooting Guides

PCR Troubleshooting Guide

Interactive Tools

Application Notes

Citations

  • Matousková, M., Vesely, P., Daniel, P., Mattiuzzo, G., Hector, R.D., Scobie, L., Takeuchi, Y. and Hejnar, J. (2013) Role of DNA methylation in expression and transmission of porcine endogenous retroviruses. PLOS ONE; PubMedID: 23986605
  • Liang, Y., Basu, D., Pattathil, S., Xu, W.L., Venetos, A., Martin, S.L., Faik, A., Hahn, M.G. and Showalter, A.M. (2013) Biochemical and physiological characterization of fut4 and fut6 mutants defective in arabinogalactan-protein fucosylation in Arabidopsis. PLOS ONE; PubMedID: 24127514
  • Megan S Thoemmes, Daniel J Fergus, Julie Urban, Michelle Trautwein, Robert R Dunn (2014) Ubiquity and diversity of human-associated demodex mites. PLoS One; 9, e106265. PubMedID: 25162399 , DOI: 10.1371/journal.pone.0106265
  • Malyshev, D.A., Dhami, K., Quach, H.T., Lavergne, T., Ordoukhanian, P., Torkamani, A. and Romesberg, F.E. (2012) Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet. Proc. Natl. Acad. Sci. USA; 109, 4503-4508. PubMedID: 22773812

Quality & Safety

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 control's 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.