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OneTaq® Hot Start DNA Polymerase
Catalog #
Size
Concentration
Price
Qty
M0481S
200 units
5,000 units/ml
$81.00
1
M0481L
1,000 units
5,000 units/ml
$324.00
1
M0481X
5,000 units
5,000 units/ml
$1,289.00
1
Categories: OneTaq® DNA Polymerases, Thermophilic DNA Polymerases
Applications: Difficult PCR, High-Fidelity PCR, High-Throughput PCR | More +
Product
Information
FAQs
Description
Properties and Usage
Protocols
Other Tools &
Resources
Related
Products
Tech Tips
Advantages and Features
Quality Control
Description
OneTaq® Hot Start DNA Polymerase is an optimized blend of Taq and Deep VentR™ DNA polymerases combined with an aptamer-based
inhibitor. The 3´→ 5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA
Polymerase (1) and the hot start formulation combines convenience with decreased interference from primer dimers and secondary
products. The OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields with minimal optimization, regardless
of a template’s GC content.
The inhibitor binds reversibly, 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. OneTaq Hot Start DNA Polymerase does not require a separate high
temperature incubation step to activate the enzyme and can be used in typical Taq-based cycling protocols.
OneTaq Hot Start 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 (i.e. ≥ 75% GC), the OneTaq High GC Enhancer can be added at a final concentration of 10–20%
to reactions containing OneTaq GC Reaction Buffer.
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 below gel. Marker M is the 1 kb DNA Ladder (NEB #N3232 ).
Highlights
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:
OneTaq® Standard Reaction Buffer
5X
OneTaq® GC Reaction Buffer
5X
OneTaq High GC Enhancer
100%
Advantages and Features
Features
Room temperature reaction setup
Applications
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
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
1.25units/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 200 µg/ml activated Calf Thymus DNA.
Quality Control
Quality Control Assays
The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data
can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
Inhibition of Primer Extension (Hot Start, Radioactivity Incorporation):
The hot start polymerase is compared to the polymerase in non-hot start conditions using primer extension; inhibition is assessed by
comparing radioactive incorporation.
PCR Amplification (Buffer Dependent, GC-rich, Master Mix) :
The polymerase master mix is tested in a polymerase chain reaction (PCR) using a GC-rich control template and specific primers,
resulting in the buffer-dependent production of the expected product.
PCR Amplification (DNA Polymerase):
The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the
expected product.
PCR Amplification (Enhancer Dependent, GC-rich, Master Mix) :
The polymerase master mix is tested in a polymerase chain reaction (PCR) using a GC-rich control template and specific primers,
resulting in the enhancer-dependent production of the expected product.
PCR Amplification (Hot Start):
The polymerase is tested in a hot start polymerase chain reaction (PCR) using Lambda DNA as the control template, specific primers
and human genomic DNA, resulting in an increase in yield of the expected Lambda product and a decrease in non-specific genomic
bands when compared to a non-hot start control reaction.
Notes
1. The OneTaq High GC Enhancer should not be used alone. It should be added only to the OneTaq GC Reaction Buffer and will typically
enhance yields when other conditions have failed.
2. Product specifications for individual components in the OneTaq Hot Start 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, 1530-1554.
3. Powell, L.M. et al. (1987). Cell. 50, 831-840.
Legal and Disclaimers
Licenses
Notice to Purchaser:
Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from
SomaLogic, Inc. (See Patent Nos. 5,475,096; 5,670,637; 5,696,249; 5,874,557; and 5,693,502). New England Biolabs, Inc. gives the
Buyer/User a non-exclusive license to use the aptamer-based OneTaq® Hot-Start DNA Polymerase for RESEARCH PURPOSES ONLY.
Commercial use of the aptamer-based OneTaq® Hot-Start DNA Polymerase requires a license from New England Biolabs, Inc. Please
contact [email protected] for more information.
The purchase of this product also conveys to the purchaser only the limited, non-transferable right to use the purchased quantity of the
product for the purchaser's own research by the purchaser only under the following U.S. patent claims and foreign counterpart patent
claims: U.S. Patent No. 5,436,149 (claims 6-16). No rights are granted to the purchaser to sell, modify for resale or otherwise transfer this
product, either alone or as a component of another product, to any third party. Takara Bio reserves all other rights, and this product may not
be used in any manner other than as provided herein. For information on obtaining a license to use this product for purposes other than
research, please contact Takara Bio Inc., Seta 3-4-1, Otsu, Shiga 520-2193, Japan (Fax +81-77-453-9254).
ONETAQ® 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.
Supporting Documents
Material Safety Datasheets
The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming
structure is used to name these document files: [Product Name]MSDS. For international versions please contact us at [email protected].
OneTaq® Hot Start DNA Polymerase MSDS
OneTaq® Standard Reaction Buffer Pack MSDS
OneTaq® GC Reaction Buffer Pack MSDS
OneTaq® High GC Enhancer MSDS
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 [email protected] or fill out the Technical Support Form for
appropriate document.
M0481S-0031112
M0481SDatasheet-Lot0051206
M0481Datasheet-Lot0051206
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How should I set up a PCR using OneTaq® Hot Start DNA Polymerase?
Can I use my regular Taq-based cycling conditions for OneTaq® DNA Polymerase based products?
Which buffer should I use?
How do I activate OneTaq® Hot Start Polymerase?
Is OneTaq® Hot Start DNA Polymerase active in other Taq reaction buffers?
When should I add the High GC Enhancer?
Can OneTaq® DNA Polymerase be used in colony PCR?
What type of DNA ends result from a primer extension reaction or a PCR using OneTaq® DNA Polymerase?
How long a product can be made by OneTaq® DNA Polymerase?
Can OneTaq® DNA Polymerase be used with uracil-containing primers or bisulfite-treated DNA?
What is the fidelity of OneTaq® DNA Polymerase?
Which buffer should I use if I want to control the level of magnesium in the reaction?
Where can I find help troubleshooting my PCR?
How should I determine an appropriate annealing temperature for my reaction?
How is OneTaq DNA Polymerase different from LongAmp Taq DNA Polymerase?
1. Protocol for OneTaq Hot Start DNA Polymerase (M0481)
Selection Tools
Troubleshooting Guides
Usage Guidelines & Tips
Interactive Tools
Selection Tools
Applications of Polymerases
DNA Polymerase Selection Chart
Thermophilic DNA Polymerases
Usage Guidelines & Tips
Guidelines for PCR Optimization with OneTaq® and OneTaq® Hot Start DNA Polymerases
Guidelines for PCR Optimization with Thermophilic DNA Polymerases
Troubleshooting Guides
PCR Troubleshooting Guide
Interactive Tools
Tm Calculator
Companion Products
Deoxynucleotide (dNTP) Solution Set
Deoxynucleotide (dNTP) Solution Mix
Diluent F
Magnesium Chloride (MgCl2) Solution
Magnesium Sulfate (MgSO4) Solution
OneTaq® (Mg-Free) GC Reaction Buffer Pack
OneTaq® (Mg-Free) Standard Reaction Buffer Pack
OneTaq® DNA Polymerase
Materials Sold Separately
OneTaq® Standard Reaction Buffer Pack
OneTaq® GC Reaction Buffer Pack
Did you know most OneTaq Hot Start reactions amplify more efficiently and robustly when you use a 68°C extension temperature?