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Thermophilic DNA Polymerases >
Deep VentR™ (exo-) DNA Polymerase |
 | | | | Deep VentR™ (exo-) DNA Polymerase | | | |
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Prices are in US dollars and valid only for US orders.
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- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer and 100 mM MgSO4
- High-Fidelity: 1.5X greater than Taq
- Extremely High Thermostability: half-life of 23 hours at 95°C
- Difficult Templates: idea for GC-rich or looped sequences
Description:
Deep VentR™ (exo-) DNA has been genetically engineered to eliminate the 3´→ 5´ proofreading exonuclease activity associated with Deep Vent DNA Polymerase. Deep Vent (exo-) DNA Polymerase is even more stable than Vent (exo-) DNA Polymerase with a half-life of 23 hours at 95°C and 8 hours at 100°C. Both Deep Vent (exo-) and Vent (exo-) DNA Polymerase are suitable for primer extensions and high temperature (72°C) DNA sequencing.
Source:
Deep VentR DNA Polymerase is purified from a strain of E. coli that carries the Deep VentR DNA Polymerase gene from Pyrococcus species GB-D. The native organism was isolated from a submarine thermal vent at 2010 meters (1) and is able to grow at temperatures as high as 104°C .
Applications:Reagents Supplied:
MgSO4 (100 mM)
ThermoPol Reaction Buffer (10X)
Enzyme Properties
Polymerase Properties | Thermophilic Polymerase Characteristics
3´ to 5´ Exonuclease: No
5´ to 3´ Exonuclease: No
Strand Displacement: Yes
Heat Inactivation:
No
Molecular Weight:
Theoretical: 89 kDa
Specific Activity:
28,000 units/mg
Reaction & Storage Conditions
Reaction Conditions:
1X ThermoPol Reaction Buffer
Supplemented with 200 μM dNTPs (not included) and 200 μg/ml activated calf thymus DNA
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
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material 30 minutes at 75°C.
Unit Assay Conditions: 1X ThermoPol Reaction Buffer, 200 µM each dNTP including [3H]-dTTP, 200 µg/ml activated calf thymus DNA.
Concentration:
2,000 units/ml
Storage Conditions:
10 mM Tris-HCl
100 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
0.1% Triton X-100
pH 7.4 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:- BSA is not provided with this enzyme since its presence is not necessary for most primer extension reactions. However, it is available free of charge, if requested when placing an order. Acetylated BSA should not be used for primer extension reactions.
- Vent Diluent is also available (NEB #B8004S). This buffer is recommended for making dilutions of Deep VentR (exo-) DNA Polymerase.
- Additional ThermoPol Reaction Buffer packs for this product are also available (NEB #B9004S). Each buffer pack contains 4 vials of 10X buffer (1.5 mls each), and 1 vial of 100 mM MgS04.
FAQs
- Can Deep Vent (exo-) DNA Polymerase be used in other buffers?
- I can't get Deep Vent (exo-) DNA Polymerase to work yet Taq DNA Polymerase works fine.
- Are the DNA fragments produced by Deep Vent (exo-) DNA Polymerase blunt-ended or do they have the single-base 3' overhang that Taq DNA Polymerase yields?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
Purified free of contaminating endonuclease and exonuclease.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 20 units of Deep VentR™ (exo-) DNA Polymerase with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (205 cpm/μg) for 4 hours at 75ºC
released < 0.1% of the total radioactivity.
Endonuclease Assay:
Incubation of a 50 μl reaction containing 20 units of Deep VentR™ (exo-) DNA Polymerase with 1 μg of ΦX174 RF I DNA for 4 hours at 75ºC or 37°C resulted in < 10% conversion to RFII as determined by agarose gel electrophoresis.
Deep Vent DNA Polymerase crystals (Ira Schildkraut and Rebecca Kucera, New England Biolabs, Inc.)
References
- Jannasch, H.W. et al. (1992) Appl. Environ. Microbiol., 58, 3472-3481.
Reagents Sold Separately
ThermoPol Reaction Buffer
Companion Products
Vent Diluent Buffer
Legal
Patents:
New England Biolabs, Inc.: U.S. Patent No. 5,352,778
New England Biolabs, Inc.: U.S. Patent No. 5,500,363
New England Biolabs, Inc.: U.S. Patent No. 5,834,285
Licenses/Patents/Disclaimers:
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.
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