Product Class: Other

Vent® (exo-) DNA Polymerase
NEBU cloned at NEB recombinant No pcr 5

Product Introduction

Vent® (exo-) DNA Polymerase offers moderate fidelity and robust performance

  • No 3´→ 5´ proofreading exonuclease activity
  • 2X higher fidelity than Taq
  • Difficult templates: ideal for GC-rich or looped sequence
  • High thermostability: half-life of 6.7 hours at 95°C for maximum activity during PCR
Catalog # Size Concentration
M0257S 200.0 units 2000 units/ml
M0257L 1000.0 units 2000 units/ml

Product Information

Description

Vent (exo-) DNA Polymerase has been genetically engineered to eliminate the 3´→ 5´ proofreading exonuclease activity associated with Vent DNA Polymerase (1). This is the preferred form for high-temperature dideoxy sequencing reactions and for high yield primer extension reactions. The fidelity of polymerization by this form is reduced to a level about 2-fold higher than that of Taq DNA Polymerase (2,3).

Product Source

An E. coli strain that carries the Vent (D141A / E143A) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase from Thermococcus litoralis (4). The native organism is capable of growth at up to 98°C and was isolated from a submarine thermal vent (5).
This product is related to the following categories:
Other PCR Polymerases Products
This product can be used in the following applications:
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

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.

Reaction Conditions

1X ThermoPol® Reaction Buffer

1X ThermoPol® Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
0.1% Triton® X-100
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Molecular Weight

Theoretical: 89000 daltons

5' - 3' Exonuclease

No

3' - 5' Exonuclease

No

Strand Displacement

+++

Resulting Ends

  • Mix of blunt and Single-base 3´ Overhangs

Unit Assay Conditions

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

Error Rate

< 190x10-6bases

Application Features

  • PCR
  • Primer extension
  • Thermal cycle sequencing
  • High temperature dideoxy-sequencing

Product Notes

  1. BSA is not provided with this enzyme since its presence is not necessary for most primer extension reactions. However, it is available as NEB #B9001. Acetylated BSA should not be used for primer extension reactions.
  2. Diluent D is also available. This diluent is recommended for making dilutions of Vent (exo-) Polymerase.
  3. Additional ThermoPol Reaction Buffer Packs for this product are also available. Each buffer pack contains 4 vials of 10X buffer (1.5 ml each) and 1 vial of 100 mM MgSO4.

References

  1. Kong, H.M., Kucera, R.B. and Jack, W.E. (1993). J. Biol. Chem.. 268, 1965-1975.
  2. Mattila, P. et al. (1991). NAR. 19, 4967-4973.
  3. Eckert, K.A. and Kunkel, T.A. et al. (1991). PCR Methods and Applications 1. 17-24.
  4. Perler, F.B. et al. (1992). PNAS USA. 89, 5577-5581.
  5. Belkin, S. and Jannasch, H.W. (1985). Arch Microbiol. 141, 181-186.

Protocols, Manuals & Usage

Protocols

  1. Guidelines for PCR Optimization for Vent DNA Polymerase
  2. Protocol for a Routine Vent (exo-) PCR

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Can Vent (exo-) DNA Polymerase be used in other buffers?
  2. Are the DNA fragments produced by Vent (exo-) DNA Polymerase blunt-ended or do they have the single-base 3' overhang that Taq DNA Polymerase yields?
  3. I can't get Vent (exo-) DNA Polymerase to work yet Taq DNA Polymerase works fine.
  4. What should I take into consideration when designing a set of PCR primers?
  5. How can I facilitate the amplification of templates with hairpin-loop structures or high GC-content?

Troubleshooting

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

Some applications in which this product can be used may be covered by patents issued and applicable in the United States and certain other countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application in which the product is used.

Trademarks

NEW ENGLAND BIOLABS®, VENT® and THERMOPOL® are registered trademarks of New England Biolabs, Inc.
DEEP VENT® is a trademark of New England Biolabs, Inc.

TRITON® is a registered trademark of Union Carbide Corporation

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