Product Class: Other

LongAmp® Hot Start Taq 2X Master Mix
cloned at NEB recombinant No hotstart pcr 5

Product Introduction

LongAmp® Hot Start Taq 2X Master Mix offers exceptional performance with long amplicons.

  • Aptamer-based inhibitor enables room temperature reaction setup
  • Optimized enzyme blend for amplification up to 30 kb
  • 2X higher fidelity than Taq
  • Master mix is a 2X concentrated solution containing everything needed for robust amplification of complex or simple templates
Catalog # Size Concentration
M0533S 100.0 reactions (50.0 µl) 2 X
M0533L 500.0 reactions (50.0 µl) 2 X

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Product Information

Description

LongAmp Hot Start Taq 2X Master Mix contains a unique blend of aptamer-based Hot Start Taq and Deep Vent® DNA Polymerases. The aptamer-based inhibitor binds reversibly to the enzyme, inhibiting polymerase activity at temperatures below 45°C, but releases the enzyme during normal PCR cycling conditions. This permits assembly of PCR reactions at room temperature. The LongAmp® Hot Start Taq 2X Master Mix does not require a separate high temperature incubation step to activate the enzyme. The 3´→5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Hot Start Taq DNA Polymerase (1). LongAmp® Hot Start Taq DNA Polymerase offers two-fold higher fidelity than Hot Start Taq DNA Polymerase alone. The convenient master mix formulation is supplied at a 2X concentration and contains dNTPs and Mg++, requiring only the addition of primers and DNA template for robust amplification. A wide range of PCR products can be generated; up to 30 kb from lambda or human genomic DNA.

Amplification of a 8 kb amplicon from varying amounts of Jurkat genomic DNA using LongAmp Hot Start Taq 2X Master Mix. Starting template amounts are indicated below the gel. Marker M is the NEB 2-Log DNA Ladder (NEB #N3200 ).

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.

Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 75°C.

Unit Assay Conditions:
1X ThermoPol™ Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 200 µg/ml activated Calf Thymus DNA.
This product is related to the following categories:
LongAmp® DNA Polymerases Products,
Master Mixes Products,
This product can be used in the following applications:
Hot Start PCR,
Long Range PCR,
Multiplex PCR,
Specialty PCR,
Routine PCR,
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

1X Master Mix Composition

60 mM Tris-SO4
20 mM (NH4)2SO4
2 mM MgSO4
3% Glycerol
0.06% IGEPAL® CA-630
0.05% Tween® 20
0.3 mM dNTPs
125 units/ml LongAmp® Hot Start Taq DNA Polymerase
pH 9.1 @ 25°C

Heat Inactivation

No

5' - 3' Exonuclease

Yes

3' - 5' Exonuclease

Yes

Strand Displacement

+

Unit Assay Conditions

1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.

Application Features

  • High-specificity Long Range PCR 
  • High-throughput PCR
  • Colony PCR

Product Notes

  1. Product specifications for individual components in the LongAmp Hot Start Taq 2X Master Mix are available separately.

References

  1. Barnes, W.M. (1994). Proc. Natl. Acad. Sci. USA. 91, 1350-1354.
  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, 372-374.
  5. Sarkar, G., Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res.. 18, 7465.

Protocols, Manuals & Usage

Protocols

  1. Protocol for LongAmp™ Hot Start Taq 2X Master Mix

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What ends will my PCR products have?
  2. What are Hot Start and WarmStart® polymerases and when would I use them?
  3. How should I determine the appropriate annealing temperature for my reaction?
  4. What are the properties of this polymerase (fidelity, product ends, max amplicon, modified base incorporation, etc.)?
  5. My results are not as expected. Where can I find troubleshooting help?
  6. Is Monarch-purified gDNA suitable for long-range PCR?

Troubleshooting

Tech Tips

Using an extension temperature of 65°C is recommended in LongAmp Hot Start Taq reactions.

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.

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. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the LongAmp® Hot Start Taq 2X Master Mix for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com. 

Trademarks

LONGAMP® 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.

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