Vent® DNA Polymerase |NEB酶试剂 New England Biolabs

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Vent® DNA Polymerase

Vent® DNA Polymerase offers moderate fidelity and robust performance

  • Possesses 3´→ 5´ proofreading exonuclease activity
  • 5X 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® DNA Polymerase |

库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
M0254V 2,000 units/ml 100 units ¥419.00
M0254S 2,000 units/ml 200 units ¥779.00
M0254L 2,000 units/ml 1,000 units ¥3,129.00
如何订购
Please enter a quantity for at least one size

产品信息

Vent® DNA Polymerase is a high-fidelity thermophilic DNA polymerase. The fidelity of Vent DNA Polymerase is 5-15-fold higher than that observed for Taq DNA Polymerase (1,2). This high fidelity derives in part from an integral 3’→5′ proofreading exonuclease activity in Vent DNA Polymerase (1,3). Greater than 90% of the polymerase activity remains following a 1 hour incubation at 95°C.


Vent® DNA Polymerase |
Amplification of Jurkat genomic DNA with Vent DNA Polymerase. Amplicon sizes are indicated below gel. Marker M is the 1 kb DNA Ladder (NEB #N3232 ).

产品来源

An E. coli strain that carries the Vent® DNA Polymerase gene 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).

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
    • M0254V     -20    
        Vent® DNA Polymerase M0254VVIAL -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
    • M0254S     -20    
        Vent® DNA Polymerase M0254SVIAL -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
    • M0254L     -20    
        Vent® DNA Polymerase M0254LVIAL -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 in 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

    分子量

    理论上的: 89000 daltons

    3′ – 5′ 核酸外切酶

    Yes

    链置换

    +

    产物末端

    • Blunt Ends

    单位活性检测条件

    1X ThermoPol Buffer, 200 µM each dNTP including [3H]-dTTP, 15 nM primed M13 DNA. 

    错配率

    < 57x10-6bases

  • 优势和特性

    应用特性

    • PCR
    • Primer extension

  • 相关产品

    相关产品

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

    单独销售的组分

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

  • 注意事项
    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. The calculated half-life of Vent DNA Polymerase at 95°C is 6.7 hours.
    3. Diluent D is also available. This buffer is recommended for making dilutions of Vent DNA Polymerase.
    4. 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.

  • 参考文献
    1. Mattila, P. et al. (1991). Nucl. Acids Res. 19, 4967-4973.
    2. Eckert, K.A. and Kunkel, T.A. (1991). PCR Methods and Applications. 1, 17-24.
    3. Kong, H.M., Kucera, R.B. and Jack, W.E. (1993). J. Biol. Chem. 268, 1965-1975.
    4. Perler, F. et al. (1992). Proc. Natl. Acad. Sci. USA. 89, 5577.
    5. Belkin, S. and Jannasch, H.W. (1985). Arch. Microbiol. 141, 181-186.

操作说明、说明书 & 用法

  • 操作说明
    1. Guidelines for PCR Optimization for Vent DNA Polymerase
    2. Protocol for a Routine Vent 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
    1. Can Vent DNA Polymerase be used in other buffers?
    2. When should Vent DNA Polymerase be used for PCR?
    3. Vent DNA Polymerase and Deep Vent DNA Polymerase produce blunt DNA ends, but my cloning depends on the primer extension product having an adenine single-base 3´ overhang.
    4. I can’t get Vent DNA Polymerase to work yet Taq DNA Polymerase works fine.
    5. Are the DNA fragments produced by Vent DNA Polymerase blunt-ended or do they have the single-base 3′ overhang that Taq DNA Polymerase yields?
    6. Can I use Vent DNA Polymerase to polish the ends of DNA fragments?
    7. I want to clone primer extension products made with Vent DNA Polymerase or Deep Vent DNA Polymerase. Are some protocols better than others?

  • 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 0.01-0.02 U/µl Vent DNA Polymerase in the reaction.

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Vent® DNA Polymerase | Powered by Bioz See more details on Bioz

质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控,以满足指定的质量标准,对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息,可从此处查阅。

  • 产品说明与变更通知

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格,这些文件的命名规则如下:[货号]_[规格]_[版本]

    • M0254S_L_v1

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控,这些文件的命名规则如下: [货号]_[规格]_[版本]_[Lot#]

    • M0254S_L_v1_0341512
    • M0254S_L_v1_0341606
    • M0254S_L_v1_0341612
    • M0254S_L_v1_0341706
    • M0254S_L_v1_0341712
    • M0254L_v1_10010829
    • M0254S_v1_10019917
    • M0254S_v1_10040202
    • M0254L_v1_10036012
    • M0254S_v1_10053543
    • M0254S_v1_10058027
    • M0254L_v1_10057546
    • M0254S_v1_10072331
    • M0254L_v1_10083499
    • M0254L_v1_10098089
    • M0254S_v1_10098090
    • M0254L_v1_10118913
    • M0254S_v1_10119649
    • M0254S_v1_10145553
    • M0254L_v1_10151854

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

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