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OneTaq® DNA Polymerases


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An optimized blend of Taq and Deep VentR™ DNA polymerases, OneTaq® and OneTaq® Hot Start DNA Polymerases offer robust amplification across a wide range of templates. The 3′–5′ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robustness of Taq, and the hot start formulation combines convenience with decreased interference from primer dimers and secondary products. Additionally, OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields with minimal optimization, regardless of a template's GC content. Both polymerases are available in master mix and Quick-Load® master mix formats.

In contrast to chemically modified or antibody-based hot start polymerases, NEB's OneTaq Hot Start utilizes aptamer technology. This unique modified oligonucelotide binds to the polymerase through non-covalent interactions, 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.

 

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OneTaq Hot Start DNA Polymerase

Amplification of a selection of sequences with varying GC content from human and C. elegans genomic DNA using OneTaq Hot Start DNA Polymerase. The presence or absence of an extended room temperature incubation does not affect performance. GC content is indicated above gel. Marker M is the 1 kb DNA Ladder (NEB# N3232).

 

OneTaq Quick-Load DNA Polymerase

For direct and fast agarose gel loading after routine PCR, such as genotyping and colony PCR, OneTaq Quick-Load DNA Polymerase is available. The product is supplied with a green 5X OneTaq Quick Load Reaction Buffer in addition to the colorless 5X OneTaq Reaction Buffer.

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.

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FAQs for OneTaq® DNA Polymerases

Protocols for OneTaq® DNA Polymerases

Advantages

  • Exceptional performance in endpoint PCR across a wide range of template
  • Robust yields with minimal optimization
  • Convenient product formats (stand-alone enzyme, master mixes, and Quick-Load® formats)
  • Hot start version allows room temperature reaction setup and does not require a separate activation step 
  • Compatible with standard Taq protocols

Applications

  • High sensitivity PCR
  • High throughput PCR
  • Routine PCR
  • GC-rich PCR
  • AT-rich PCR
  • Primer extension
  • Colony PCR
  • Long PCR (up to ~6 kb genomic)

DNA Polymerase Selection Chart

NEB offers a guidelines for choosing the correct DNA polymerase for your application by providing a list of specific properites.
Several factors govern which polymerase should be used in a given application, including: 

Template/product specificity: Is RNA or DNA involved? Is the 3´ terminus at a gap, nick or at the end of the template? 

Removal of existing nucleotides: Will the nucleotide(s) be removed from the existing polynucleotide chain as part of the protocol? If so, will they be removed from the 5´ or the 3´ end? 

Thermal stability: Does the polymerase need to survive incubation at high temperature or is heat inactivation desirable? 

Fidelity: Will subsequent sequence analysis or expression depend on the fidelity of the synthesized products?

OneTaq 2X Master Mix with GC Buffer

Amplification of a selection of sequences with varying GC content from human genomic DNA using OneTaq 2X Master Mix with GC Buffer. GC content is indicated above gel. Marker M is the 1 kb DNA Ladder (NEB# N3232).

OneTaq 2X Master Mix with Standard Buffer

Amplification of a selection of sequences with varying GC content from human and C. elegans genomic DNA using OneTaq 2X Master Mix with Standard Buffer. Amplicon sizes are indicated next to gel, and GC content is indicated above gel. Marker M is the 1 kb DNA Ladder (NEB# N3232).

OneTaq Buffer Recommendations

NEB recommends choosing your OneTaq Buffer based on amplicon % GC content.

OneTaq Hot Start vs Other Commercially Available Hot Start Polymerases

Comparison of OneTaq Hot Start DNA Polymerases to Other Commercially Available Hot Start Polymerases
Reactions containing high GC human genomic DNA templates were set up at room temperature. PCR experiments included 30 cycles. Purity (A) and Yield (B) were calculated via microfluidic analysis from triplicate reactions. OneTaq polymerases were used with GC Buffer. Some OneTaq reactions also contained High GC Enhancer (striped bars). Competitor polymerases were cycled according to manufacturer's recommendations and included GC enhancers when supplied (striped bars).

OneTaq vs Other Commercially Available Polymerases

Comparison of OneTaq DNA Polymerase to Other Commercially Available Polymerases (non-hot start)
Amplification of a selection of high GC human genomic DNA templates demonstrates OneTaq performance. PCR experiments included 30 amplification cycles. Purity (A) and Yield (B) were calculated via microfluidic analysis from triplicate reactions. Competitor polymerases were cycled according to manufacturer's recommendations.

PCR Selection Tool

Choose from one of the largest selections of polymerases for PCR applications from the leader in enzyme technology and bring unparalleled confidence to your experiments.

Recommended Time for Enzyme Activation

Recommended time for enzyme activation of commercially available Hot Start