Vent DNA polymerase is a high-fidelity, thermostable DNA polymerase. The fidelity of Vent DNA polymerase is 5-15 times higher than that of Taq DNA polymerase (1,2). This high fidelity is due in part to the intact 3´→5´ exonuclease proofreading activity of Vent DNA polymerase (1,3). After incubation at 95°C for 1 hour, >90% of the polymerase activity is retained.
Product Source
Escherichia coli strain carrying the Vent® DNA polymerase gene cloned from Thermococcus litoralis (4). This protist was isolated from a submarine volcanic vent (5) and can grow at temperatures up to 98°C.
Characteristics and Usage
Unit Definition
One unit is the amount of enzyme required to incorporate 10 nmol of dNTPs into acid-insoluble material at 75°C in 30 minutes.
Reaction Conditions
1X ThermoPol® Reaction Buffer Set
1X ThermoPol® Reaction Buffer Set
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 Solution
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
Unit Activity Assay Conditions
1X ThermoPol Buffer, 200 µM each dNTP (including [⁺]-dTTP), 15 nM primer-bound M13 DNA.
Error Rate
< 57x10-6 bases
Advantages and Features
Applications
PCR
Primer Extension
Notes
1. This enzyme is not supplied with BSA, as most primer extension reactions do not require it. However, if required, NEB #B9001 is available. Acetylated BSA cannot be used in primer extension reactions.
2. The half-life of Deep Vent DNA Polymerase at 95°C is 6.7 hours.
3. Diluent D is available. This buffer is recommended for diluting Vent DNA Polymerase.
4. Other ThermoPol Reaction Buffer Sets are also available. Each set contains four tubes of 10X buffer (1.5 ml/tube) and one tube of 100 mM MgSO4.
Vent DNA 聚合酶是一种高保真耐热 DNA 聚合酶。Vent DNA 聚合酶的保真度比 Taq DNA 聚合酶的保真度高 5 - 15 倍（1,2）。这种高保真度部分源自 Vent DNA 聚合酶中完整的 3´→5´ 核酸外切酶校读活性（1,3）。在 95℃ 条件下温育 1 小时后，仍保留 90% 以上的聚合酶活性。
产品来源
大肠杆菌菌株，携带有克隆自热球菌（Thermococcus litoralis）（4）的 Vent® DNA 聚合酶基因。这种原生生物从海底火山口（5）中分离出来，可在高达 98℃ 的温度下生长。
特性和用法
单位定义
1 单位是指在 75℃ 条件下，30 分钟能使 10 nmol 的 dNTP 掺入酸不溶物所需的酶量。
反应条件
1X ThermoPol® 反应缓冲液套装
1X ThermoPol® 反应缓冲液套装
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
单位活性检测条件
1X ThermoPol 缓冲液、每种浓度为 200 µM 的 dNTP（含 [3H]-dTTP）、15 nM 已结合引物的 M13 DNA。
错配率
< 57x10-6bases
优势和特性
应用特性
PCR
引物延伸
注意事项
1. 该酶不提供 BSA，因为大多数引物延伸反应不需要 BSA。但如果需要，在售有 NEB #B9001。乙酰化的 BSA 不能用于引物延伸反应。
2. Deep Vent DNA 聚合酶在 95℃ 的半衰期为 6.7 小时。
3. 在售稀释液 D。推荐使用此缓冲液稀释 VentDNA 聚合酶。
4. 本产品还提供其它 ThermoPol 反应缓冲液套装。每种缓冲液套装包含 4 管 10X 缓冲液（1.5 ml/管）和 1 管 MgSO4（100 mM）。