Hi-T7® RNA Polymerase |NEB酶试剂 New England Biolabs

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产品信息

Hi-T7 RNA Polymerase is an engineered DNA-dependent RNA polymerase that is highly specific for T7 phage promoters and is designed to function at higher temperatures than the wild-type bacteriophage T7 RNA Polymerase. Hi-T7 RNA Polymerase functions at an optimal temperature of 50-52°C.

产品来源

An E. coli strain that carries an engineered His-tag T7 RNA Polyermase gene.

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

  • 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
    • M0658S     -20    
        Hi-T7® RNA Polymerase M0658SVIAL -20 1 x 0.1 ml 50,000 units/ml
        Hi-T7® RNA Polymerase Reaction Buffer B0658AVIAL -20 1 x 1 ml Not Applicable

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to incorporate 1 nmol of ATP into acid-insoluble material in 1 hour at 50°C.

    Reaction Conditions
    1X Hi-T7 RNA Polymerase Reaction Buffer, supplemented with 0.5 mM each of ATP, UTP, GTP, CTP, and DNA template containing the T7 RNA Polymerase promoter.  Incubate at 37-56°C for 1 hour.  Optimal incubation temperature is 50-52°C.

    反应条件

    1X Hi-T7® RNA Polymerase Reaction Buffer
    Supplement with 0.5 mM ATP, 0.5 mM UTP , 0.5 mM GTP 和 0.5 mM CTP 
    Incubate at 50°C

    1X Hi-T7® RNA Polymerase Reaction Buffer
    40 mM Tris-HCl
    4 mM MgCl2
    50 mM NaCl
    2 mM spermidine
    1 mM DTT
    (pH 7.5 @ 25°C)

    贮存溶液

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

    单位活性检测条件

    1X Hi-T7 RNA Polymerase Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP and 1 µg T7 phage DNA in 50 µl reaction volume.

  • 优势和特性

    应用特性

    • Radiolabeled RNA probes
    • Non-isotopic RNA labeling
    • Guide RNA for gene targeting
    • mRNA for in vitro translation, microinjection and transfection 
    • RNA structure, processing and catalysis studies
    • RNA amplification
    • Isothermal amplification
    • Anti-sense RNA for gene expression

  • 相关产品

    相关产品

    • m0251-t7-rna-polymerase
    • 小鼠 RNase 抑制剂
    • rNTP 套装
    • rNTP 混合液

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for Standard RNA Synthesis with Hi-T7 RNA Polymerase (NEB #M0658)

  • 应用实例

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

FAQs & 问题解决指南

  • FAQs

    1. Can I use a different reaction buffer for in vitro transcription with Hi-T7 RNA Polymerase?
    2. Does Hi-T7 RNA Polymerase recognize the same promoter sequence as the wild-type T7 RNA Polymerase from bacteriophage?
    3. At what temperature is Hi-T7 RNA Polymerase active?
    4. What are the main causes of reaction failure when using Hi-T7 RNA Polymerase?
    5. What templates can be used for in vitro transcription with Hi-T7 RNA Polymerase?
    6. Can Hi-T7 RNA Polymerase transcribe off of an uncut plasmid?