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Deep Vent® (exo-) DNA Polymerase
Deep Vent® (exo-) offers moderate fidelity and robust performance
Extremely high thermostability: half-life of 23 hours at 95°C
5X higher fidelity than Taq
Difficult templates: ideal for GC-rich or looped sequence
产品信息
Deep Vent (exo-) DNA Polymerase 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.
产品来源
An E. coli strain that carries the Deep Vent (D141A / E143A) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase 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.
产品类别:
Other PCR Polymerases
应用:
Routine PCR,
PCR
产品组分信息
产品组分信息
本产品提供以下试剂或组分:
NEB #
名称
组分货号
储存温度
数量
浓度
M0259V
-20
Deep Vent® (exo-) DNA Polymerase
M0259VVIAL
-20
1 x 0.05 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
M0259S
-20
Deep Vent® (exo-) DNA Polymerase
M0259SVIAL
-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
M0259L
-20
Deep Vent® (exo-) DNA Polymerase
M0259LVIAL
-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
特性和用法
单位定义
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.
反应条件
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)
贮存溶液
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
热失活
否
分子量
理论上的: 89000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
No
链置换
+++
产物末端
Mix of blunt and Single-base 3´ Overhangs
单位活性检测条件
1X ThermoPol Reaction Buffer, 200 µM each dNTP including [3H]-dTTP, 15 nM primed M13 DNA.
优势和特性
应用特性
PCR
Primer extension
相关产品
相关产品
Deoxynucleotide (dNTP) Solution Mix
Deoxynucleotide (dNTP) Solution Set
单独销售的组分
ThermoPol® Reaction Buffer Pack
Magnesium Sulfate (MgSO4) Solution
注意事项
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 (#B8004). This diluent is recommended for making dilutions of Deep Vent (exo-) DNA Polymerase.
Additional ThermoPol Reaction Buffer Packs are also available (#B9004). Each buffer pack contains 4 vials of 10X buffer (1.5 ml each) and 1 vial of 100 mM MgSO4.
参考文献
Jannasch, H.W. et al. (1992). Appl. Environ. Mircobiol. 58, 3472-3481.
操作说明、说明书 & 用法
操作说明
Guidelines for PCR Optimization for Deep Vent® DNA Polymerase
Protocol for a Routine Deep Vent® (exo-) PCR
使用指南
Guidelines for PCR Optimization with Thermophilic DNA Polymerases
工具 & 资源
选择指南
DNA Polymerase Selection Chart
Thermophilic DNA Polymerases
FAQs & 问题解决指南
FAQs
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?
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.
问题解决指南
PCR Troubleshooting Guide
实验技巧
The amount of enzyme in the reaction is critical. Try a final concentration of 0.02-0.04 U/µl Deep Vent (exo-) DNA Polymerase in the reaction.