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Thermophilic DNA Polymerases >
Deep VentR™ DNA Polymerase |
 | | | | Deep 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
- Extremely High Thermostability: half-life of 23 hours at 95°C
- Difficult Templates: ideal for GC-rich or looped sequences
Description:
Deep VentR™ DNA Polymerase is the second high-fidelity thermophilic DNA polymerase available from New England Biolabs. The fidelity of Deep VentR DNA Polymerase is derived in part from an integral 3´→ 5´ proofreading exonuclease activity. Deep VentR is even more stable than VentR at temperatures of 95 to 100°C.
Amplification of Jurkat genomic DNA with Deep Vent DNA Polymerase. Amplicon sizes are indicated below gel. Marker M is the 1 kb DNA Ladder (NEB #3232).
Source:
An E. coli strain that carries the Deep Vent DNA Polymerase gene from Pyrococcus species GB-D. The native organism was isolated from a submarine thermal vent at 2,010 meters (1) and is able to grow at temperatures as high as 104°C.
Applications:Reagents Supplied:
MgSO4 (100 mM)
ThermoPol Reaction Buffer Pack (10X)
Enzyme Properties
Polymerase Properties | Thermophilic Polymerase Characteristics
3´ to 5´ Exonuclease: Yes
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 Pack
Supplemented with 200 μM dNTPs (not included) and 200 μg/ml activated calf thymus DNA
1X ThermoPol Reaction Buffer Pack:
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 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:- The calculated half-life of Deep VentR DNA Polymerase at 95°C is 23 hours.
- Vent Diluent is also available (NEB# B8004S). This buffer is recommended for making dilutions of Deep VentR 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.
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 Deep Vent DNA Polymerase be used in other buffers?
- I can't get Deep Vent DNA Polymerase to work yet Taq DNA Polymerase works fine.
- Are the DNA fragments produced by Deep Vent DNA Polymerase blunt-ended or do they have the single-base 3' overhang that Taq DNA Polymerase yields?
- 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.
- Can I use Deep 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?
Protocols
Protocols for Deep VentR™ DNA Polymerase
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 in ThermoPol Reaction Buffer supplemented with 400 μM each dNTP containing a minimum of 20 units of Deep VentR DNA Polymerase with 1 μg of supercoiled ΦX174 DNA for 4 hours at 37°C results in < 10% conversion to the nicked form as determined by agarose gel electrophoresis.
Physical Purity:
Purified to > 95% homogeneity as determined by SDS-PAGE analysis using Coomassie Blue detection.
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 Pack
Companion Products
BSA
Deoxynucleotide Solution Mix
Deoxynucleotide Solution Set
Diluent D
Magnesium Sulfate (MgSO4) Solution
ThermoPol DF (Detergent-free) Reaction Buffer Pack
ThermoPol II (Mg-free) Reaction Buffer Pack
ThermoPol Reaction Buffer Pack
Legal
Patents:
New England Biolabs, Inc.: 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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