T7 RNA Polymerase |NEB酶试剂 New England Biolabs

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产品信息

Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 KD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology. 

Reaction Conditions
1X RNAPol Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP, and DNA template containing the T7 RNA Polymerase promoter. Incubate at 37°C. 

产品来源

Isolated from E.coli carrying a plasmid which contains the T7 RNA Polymerase gene. 

产品类别:
RNA Synthesis In vitro Transcription (IVT)

应用:
Transcription-Mediated and NASBA Amplification

  • 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
    • M0251S     -20    
        T7 RNA Polymerase M0251SVIAL -20 1 x 0.1 ml 50,000 units/ml
        RNAPol Reaction Buffer B9012SVIAL -20 1 x 1 ml 10 X
    • M0251L     -20    
        T7 RNA Polymerase M0251LVIAL -20 1 x 0.5 ml 50,000 units/ml
        RNAPol Reaction Buffer B9012SVIAL -20 1 x 1 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme that will incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50 μl in 1 hour at 37°C.

    反应条件

    1X RNAPol Reaction Buffer
    Incubate at 37°C

    1X RNAPol Reaction Buffer
    40 mM Tris-HCl
    6 mM MgCl2
    1 mM DTT
    2 mM spermidine
    (pH 7.9 @ 25°C)

    贮存溶液

    50 mM Tris-HCl
    100 mM NaCl
    20 mM β-ME
    1 mM EDTA
    50% Glycerol
    0.1% (w/v) Triton® X-100
    pH 7.9 @ 25°C

    单位活性检测条件

    1X RNAPol Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP, and 1 µg T7 DNA in 50 µl.

  • 优势和特性

    应用特性

    • Radiolabeled RNA probes
    • Non-isotopic RNA labeling
    • Preparation of RNA vaccines
    • Guide RNA for gene targeting
    • mRNA for in vitro translation and micro injection 
    • RNA structure, processing and catalysis studies
    • RNA amplification
    • Anti-sense RNA for gene expression experiment 

  • 相关产品

    相关产品

    • e2040-hiscribe-t7-high-yield-rna-synthesis-kit
    • ssRNA Ladder
    • 小鼠 RNase 抑制剂
    • 无机焦磷酸酶（大肠杆菌）
    • 2X RNA 上样染料
    • E. coli Poly(A) 聚合酶
    • mRNA 帽结构 2-O-甲基转移酶
    • m2080-vaccinia-capping-system

  • 注意事项

    1. For radio labeled high specific activity RNA probes, the concentration of the radioactive nucleotide should be limited to 6 μM. 
    2. To protect RNA against ribonuclease, RNase inhibitor (NEB #M0314 or #M0307) should be added to a final concentration of 1 U/ μl. 
    3. T7 RNA Polymerase is extremely sensitive to salt inhibition. The overall salt concentration should not exceed 50 mM. 

  • 参考文献

    1. Schenborn, E.T. and Meirendorf, R.C. (1985). Nucl. Acids Res.. 13, 6223-6236.
    2. Davanloo, P., Rosenberg, A.H., Dunn, J.J. and Studier, F.W. (1984). Proc. Natl. Acad. Sci. USA. 81, 2035-2039.
    3. Sambrook, J., Fritsch, E.F. and Maniatis, T. Molecular Cloning: A Laboratory Manual, (2nd Ed.). 10.27-10.37.
    4. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, (2nd Ed.). 18.82-18.84.
    5. Melton, D.A., Kreig, P.A., Rebagliati, M.R., Maniatis, T., Zinn, K. and Green, M.R. (1984). Nucl. Acids Res.. 12, 7035-7056.
    6. Milligan, J.F., Groebe, D.R., Witherell, G.W. and Uhlenbeck, O.C. (1987). Nucl. Acids Res.. 15, 8783.
    7. Noren, C.J. et al. (1990). Nucl. Acids Res.. 18, 83-88.
    8. Kreig, P.A. and Melton, D.A. (1984). Nucl. Acids Res.. 12, 7057-7070.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for Standard RNA Synthesis

  • 应用实例

    • Scaling of High-Yield In vitro Transcription Reactions for Linear Increase of RNA Production

工具 & 资源

  • 选择指南

    • DNA Polymerase Selection Chart
    • RNA Polymerase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. What is the promoter sequence of T7 RNA Polymerase?
    2. Is it possible to start transcription with an A?
    3. Does the transcription reaction with T7 RNA Polymerase require a primer?
    4. Does T7 RNA Polymerase leave an extra base at the end of a transcript?
    5. Will T7 RNA Polymerase work on single stranded substrate?
    6. Will T7 RNA Polymerase work on uncut plasmid DNA?
    7. Can aberrant RNA be produced when using T7 RNA Polymerase?
    8. How can the yield of RNA be maximized when using T7 RNA Polymerase?
    9. Can I use T7 RNA Polymerase to make high specific activity radiolabeled probes?
    10. What are the main causes of reaction failure using T7 RNA Polymerase?
    11. Why is the specific activity of the probe low?
    12. How can the yield of RNA be maximized when using T7 RNA Polymerase?