Vent® (exo-) DNA Polymerase

Vent® (exo-) DNA Polymerase

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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

: BP-0004
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CHI TIẾT :

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).

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

  • M0257S

     

     

    -20

     

     

     

    Vent® (exo-) DNA Polymerase

    M0257SVIAL

    -20

    1 x 0.1 ml

    2,000 units/ml

     

    ThermoPol® Reaction Buffer Pack

    B9004SVIAL

    -20

    1 x 1.5 ml

    10 X

     

    Magnesium Sulfate (MgSO4) Solution

    B1003SVIAL

    -20

    1 x 1.5 ml

    100 mM

  • M0257L

     

     

    -20

     

     

     

    Vent® (exo-) DNA Polymerase

    M0257LVIAL

    -20

    1 x 0.5 ml

    2,000 units/ml

     

    ThermoPol® Reaction Buffer Pack

    B9004SVIAL

    -20

    3 x 1.5 ml

    10 X

     

    Magnesium Sulfate (MgSO4) Solution

    B1003SVIAL

    -20

    1 x 1.5 ml

    100 mM

Advantages and Features

Application Features

  • PCR

  • Primer extension

  • Thermal cycle sequencing

  • High temperature dideoxy-sequencing

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 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)

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

Related Products

Companion Products

  • Deoxynucleotide (dNTP) Solution Set
  • Deoxynucleotide (dNTP) Solution Mix

Materials Sold Separately

  • ThermoPol® Reaction Buffer Pack
  • Magnesium Sulfate (MgSO4) Solution

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.