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产品中心 酶制剂 分子酶

T4 DNA Ligase

价格 ￥200.00 （ 1-2周）
货号 UA070112
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产品介绍

产品介绍 评论(0)

产品规格

  • 浓度

    5 U/μL

  • 表达宿主

    E.coli

  • 分子量

    55.3 kDa

  • 纯度

    ＞95% by SDS-PAGE

  • 性状

    Liquid

  • 缓冲体系

    Storage Buffer: 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA and 50% glycerol.

    1 × Reaction Buffer: 40 mM Tris-HCl (pH 7.8 @ 25°C), 10 mM MgCl2, 10 mM DTT, 0.5 mM ATP.


  • 储存条件

    -20℃

背景介绍

  • T4 DNA ligase catalyzes the formation of phosphodiester bonds between adjacent 5'-phosphate and 3'-hydroxyl ends in double-stranded DNA or RNA. This enzyme can repair single-strand nicks in double-stranded DNA, RNA, or DNA/RNA hybrids. It can also join DNA fragments with sticky or blunt ends, but is inactive toward single-stranded nucleic acids.
    T4 DNA ligase requires ATP as a cofactor.

    Application:

    Cloning of restriction enzyme generated DNA fragments.

    Cloning of PCR products.

    Joining of double-stranded oligonucleotide linkers or adaptors to DNA.

    Site-directed mutagenesis.

    Amplified fragment length polymorphism (AFLP).

    Ligase-mediated RNA detection.

    Nick repair in duplex DNA, RNA or DNA/RNA hybrids.

    Self-circularization of linear DNA

操作步骤

  • Reaction Conditions: 1×T4 DNA Ligase Reaction Buffer. Incubate at 16 °C.

    Inhibition and Inactivation: T4 DNA Ligase is strongly inhibited by NaCl or KCl at concentrations more than 200 mM.

    Inactivated by heating at 65 °C for 10 min or at 70 °C for 5 min.

酶活定义
  • One Weiss unit of the enzyme catalyzes the conversion of 1 nmol of [32PPi] into Norit-adsorbable form in 20 min at 37°C. Enzyme activity is assayed in the following mixture: 66 mM Tris-HCl (pH 7.6), 6.6 mM MgCl2, 0.066 mM ATP, 10 mM DTT, 3.3 µM [32PPi]. 1 Weiss Unit = 200 CEU One CEU is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of lambda DNA in 30 min at 16°C.

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