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OneTaq® DNA Polymerase

  The ONE You've Been Waiting For

The OneTaq® contest is now over. Thank you to everyone who entered, and congratulations to our winners! Click here to see what others are saying about OneTaq.  



"... OneTaq worked like a charm"
"I am very happy with the performance of this polymerase. We've been having quite some trouble with genotyping, and OneTaq worked like a charm."
– Dubravka Pezic
Postdoctoral Scholar
California Institute of Technology

"The OneTaq polymerase worked wonders!..."
"The OneTaq polymerase worked wonders! I tried multiple conditions with other commercial polymerase for my PCR reactions and resulted in low yield. When I tried OneTaq polymerase, the result was an instant gratification!"

– Hua Wang
Graduate Student
University of Oxford

"... OneTaq basically blew away the competition."
"We were experiencing problems amplifying this particular molecule after our capture experiment. OneTaq basically blew away the competition."

– Chun-Nan Chen, Ph.D.
Chief Scientific Officer
Single Cell Technology, Inc.

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 primerdimers 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.


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)


Achieve robust amplification for routine, AT- and GC-rich
templates with OneTaq


Amplification of a selection of sequences with varying AT and 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.



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.


OneTaq Buffer Recommendations
AMPLICON %
GC CONTENT
RECOMMENDED
DEFAULT BUFFER
OPTIMIZATION NOTES
<50% GC OneTaq Standard Reaction Buffer* Adjust annealing temperature, primer/
template concentration, etc., if needed.
50–65% GC OneTaq Standard Reaction Buffer* OneTaq GC Reaction Buffer can be
used to enhance performance of difficult
amplicons.
>65% GC OneTaq GC Reaction Buffer* OneTaq GC Reaction Buffer with
10–20% OneTaq High GC Enhancer
can be used to enhance performance of
difficult amplicons.
* Mg-free formulations are also available.


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
.



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.
  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.


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).


Table 1: Recommended time for enzyme activation of commercially available Hot Start Taq products
MANUFACTURER ENZYME ACTIVATION STEP* HOT START FORM
Applied BioSystems AmpliTaq Gold® 360 10', 95°C Modified
Invitrogen Platinum® Taq 30"–2', 94°C Ab
Promega GoTaq® Hot Start 2', 94–95°C Ab
Qiagen HotStarTaq 15', 95°C Modified
Roche FastStart Taq 4', 95°C Modified
Sigma JumpStart™ Taq 1', 94°C Ab
Thermo Fisher Thermo-Start Taq 15', 95°C Modified
NEB OneTaq None Aptamer
* May include initial denaturation step.


Ordering Information

PRODUCT NEB # SIZE
OneTaq® DNA Polymerase M0480S/L/X 200/1,000/5,000 units
OneTaq® 2X Master Mix with Standard Buffer M0482S/L 100/500 rxns (50μl vol)
OneTaq® 2X Master Mix with GC Buffer M0483S/L 100/500 rxns (50μl vol)
OneTaq® Quick-Load® 2X Master Mix
with Standard Buffer
M0486S/L 100/500 rxns (50μl vol)
OneTaq® Quick-Load® 2X Master Mix
with GC Buffer
M0487S/L 100/500 rxns (50μl vol)
OneTaq® Hot Start DNA Polymerase M0481S/L/X 200/1,000/5,000 units
OneTaq® Hot Start 2X Master Mix with Standard Buffer M0484S/L 100/500 rxns (50μl vol)
OneTaq® Hot Start 2X Master Mix with GC Buffer M0485S/L 100/500 rxns (50μl vol)
OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer M0488S/L 100/500 rxns (50μl vol)
OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer M0489S/L 100/500 rxns (50μl vol)
COMPANION PRODUCTS
OneTaq® Standard Reaction Buffer Pack B9022S 8 ml (4 x 2.0 ml)
OneTaq® GC Reaction Buffer Pack B9023S 8 ml (4 x 2.0 ml)
OneTaq® (Mg-Free) Standard Reaction Buffer Pack B9024S 8 ml (4 x 2.0 ml)
OneTaq® (Mg-Free) GC Reaction Buffer Pack B9025S 8 ml (4 x 2.0 ml)


AmpliTaq Gold® is a registered trademark of Life Technologies.
Platinum® is a registered trademark of Invitrogen.
GoTaq® is a registered trademark of Promega.
JumpStart™ is a trademark of Sigma.