T4 RNA Ligase 2 (dsRNA Ligase) |NEB酶试剂 New England Biolabs

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产品信息

T4 RNA Ligase 2, also known as T4 Rnl2 (gp24.1), has both intermolecular and intramolecular RNA strand joining activity (1-3). Unlike T4 RNA Ligase 1 (NEB #M0204), T4 RNA Ligase 2 is much more active joining nicks on double stranded RNA than on joining the ends of single stranded RNA. The enzyme requires an adjacent 5´ phosphate and 3´ OH for ligation. The enzyme can also ligate the 3´ OH of RNA to the 5´ phosphate of DNA in a double stranded structure (4).

产品来源

An E. coli strain that carries the T4 RNA Ligase 2 gene.

产品类别:
RNA Ligation

  • 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
    • M0239S     -20    
        T4 RNA Ligase 2 (dsRNA Ligase) M0239SVIAL -20 1 x 0.015 ml 10,000 units/ml
        T4 Rnl2 Reaction Buffer B0239SVIAL -20 1 x 1.5 ml 10 X
    • M0239L     -20    
        T4 RNA Ligase 2 (dsRNA Ligase) M0239LVIAL -20 1 x 0.075 ml 10,000 units/ml
        T4 Rnl2 Reaction Buffer B0239SVIAL -20 1 x 1.5 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to ligate 0.4 µg of an equimolar mix of a 23-mer and 17-mer RNAs in a total reaction volume of 20 µl in 30 minutes at 37°C.

    反应条件

    1X T4 Rnl2 Reaction Buffer
    Incubate at 37°C

    1X T4 Rnl2 Reaction Buffer
    50 mM Tris-HCl
    2 mM MgCl2
    1 mM DTT
    400 µM ATP
    (pH 7.5 @ 25°C)

    贮存溶液

    10 mM Tris-HCl
    50 mM KCl
    35 mM (NH4)2SO4
    0.1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    pH 7.5 @ 25°C

    热失活

    80°C for 5 minutes

    比活度

    40,000 units/mg

    单位活性检测条件

    1X T4 Rnl2 Reaction Buffer and 0.4 μg of an equimolar mix of the 23-mer and 17-mer RNAs. After incubation at 37°C for 30 minutes, the ligated product is detected on a 15% polyacrylamide gel.

  • 优势和特性

    应用特性

    • Ligate a nick in dsRNA
    • Ligate the 3´ OH of RNA to the 5´ phosphate of DNA in a double stranded structure

  • 相关产品

    相关产品

    • m0204-t4-rna-ligase-1-ssrna-ligase
    • 小鼠 RNase 抑制剂

  • 参考文献

    1. Ho, C.K. and Shuman, S. (2002). Proc. Natl. Acad. Sci. USA. 99, 12709-12714.
    2. Ho, C.K. et al. (2004). Structure. 12, 327-339.
    3. Nandakumar, J. et al. (2004). J. Biol. Chem.. 279, 31337-31347.
    4. Nandakuman, J. and Shuman, S. (2004). Mol. Cell. 16, 211-221.

操作说明、说明书 & 用法

  • 操作说明

    1. Protocol for ligation of Nicks in dsRNA with T4 RNA Ligase 2 (NEB #M0239)
    2. Protocol for ligation of 3′OH of RNA and 5′PO4 of DNA in ds structure with T4 RNA Ligase 2 (NEB #M0239)

工具 & 资源

  • 选择指南

    • Properties of DNA and RNA Ligases
    • RNA Ligase Selection Chart
    • Substrate-based Ligase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. How can I achieve optimal ligation of a 5’ phosphorylated ssDNA oligo to the 3’ end of an ssRNA substrate with a ssDNA or ssRNA splint using T4 RNA Ligase 2?