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VentR® 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: 5X greater than Taq
- High Thermostability: half-life of 6.7 hours at 95°C
- Value: for routine high-fidelity PCR
Description:
VentR® DNA Polymerase is a high-fidelity thermophilic DNA polymerase. The fidelity of VentR DNA Polymerase is 5-15-fold higher than that observed for Taq DNA Polymerase (1,2). This high fidelity derives in part from an integral 3'→5' proofreading exonuclease activity in VentR DNA Polymerase (1,3). Greater than 90% of the polymerase activity remains following a 1 hour incubation at 95°C.
Source:
Both VentR and VentR (exo-) are purified from strains of E. coli that carry the Vent DNA Polymerase gene from the archaea Thermococcus litoralis (4). The native organism is capable of growth at up to 98°C and was isolated from a submarine thermal vent (5).
Applications:Reagents Supplied:
MgSO4 (100 mM)
ThermoPol Reaction Buffer (10X)
Enzyme Properties
Polymerase Properties | Thermophilic Polymerase Characteristics
3´ to 5´ Exonuclease: Yes
5´ to 3´ Exonuclease: No
Strand Displacement: Yes
Error Rate: 57
x10-6 bases
Heat Inactivation:
No
Molecular Weight:
Theoretical: 89,000 daltons
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 in a total reaction volume of 50 μl in 30 minutes at 75°C.
Unit Assay Conditions: 1X ThermoPol 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
The calculated half-life of VentR DNA Polymerase at 95°C is 6.7 hours.General notes:- Vent Diluent is also available (NEB #B8004). This buffer is recommended for making dilutions of VentR DNA Polymerase.
- Additional buffer packs for this product are also available (NEB #B9004S). Each buffer pack contains 4 vials of 10X buffer (1.5 ml each), and 1 vial of 100 mM MgS04.
Usage 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.
FAQs
- Can Vent DNA Polymerase be used in other buffers?
- When should Vent DNA Polymerase be used for PCR?
- Vent DNA Polymerase (NEB #M0254) and Deep Vent DNA Polymerase (NEB #M0258) produce blunt DNA ends, but my cloning depends on the primer extension product having an adenine single-base 3´ overhang.
- I can't get Vent DNA Polymerase to work yet Taq DNA Polymerase works fine.
- Are the DNA fragments produced by Vent DNA Polymerase blunt-ended or do they have the single-base 3' overhang that Taq DNA Polymerase yields?
- Can I use Vent DNA Polymerase to polish the ends of DNA fragments?
- I want to clone primer extension products made with Vent DNA Polymerase or Deep Vent DNA Polymerase. Are some protocols better than others?
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 endonucleases and exonucleases.
Endonuclease Assay:
Incubation of a 50 μl reaction containing 20 units of VentR® DNA Polymerase with 1 μg of pUC19 DNA in 1X assay buffer supplemented with 400 µM each dNTP for 4 hours at 37°C resulted in <10% conversion to RFII as determined by agarose gel electrophoresis.
References
- Mattila, P. et al. (1991) Nucl. Acids Res., 19, 4967-4973.
- Eckert, K.A. and Kunkel, T.A. (1991) PCR Methods and Applications, 1, 17-24.
- Kong, H.M., Kucera, R.B. and Jack, W.E. (1993) J. Biol. Chem., 268, 1965-1975.
- Perler, F. et al. (1992) Proc. Natl. Acad. Sci. USA, 89, 5577.
- Belkin, S. and Jannasch, H.W. (1985) Arch. Microbiol., 141, 181-186.
Reagents Sold Separately
ThermoPol Reaction Buffer
Companion Products
Vent Diluent Buffer
Legal
Patents:
New England Biolabs: U.S. Patent No. 5,210,036
New England Biolabs: U.S. Patent No. 5,500,363
New England Biolabs: U.S. Patent No. 5,352,778
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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