Brand: New England Biolabs (UK) Ltd
Vent® 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
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).
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).
The following reagents are supplied with this product:
NEB # |
Component Name |
Component # |
Stored at (°C) |
Amount |
Concentration |
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PCR
Primer extension
Thermal cycle sequencing
High temperature dideoxy-sequencing
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.
1X ThermoPol® Reaction Buffer Pack
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)
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
No
Theoretical: 89000 daltons
No
No
Mix of blunt and Single-base 3´ Overhangs
1X ThermoPol Reaction Buffer, 200 µM each dNTP including [3H]-dTTP, 15 nM primed M13 DNA.
< 190x10-6bases
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.
Diluent D is also available. This diluent is recommended for making dilutions of Vent (exo-) Polymerase.
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.