T4 RNA Ligase 1 (ssRNA Ligase), High Concentration |NEB酶试剂 New England Biolabs

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产品信息

T4 RNA Ligase 1, High Concentration (M0437M), is a 3X higher concentration than our standard T4 RNA Ligase 1 (M0204S/L). This product is packaged with a vial of 100 mM ATP (N0451A). The increased concentration of reaction components is helpful in lower volume reactions where reagent volumes are limited.

T4 RNA Ligase 1 catalyzes the ligation of a 5´ phosphoryl-terminated nucleic acid donor to a 3´ hydroxyl-terminated nucleic acceptor through the formation of a 3´ → 5´ phosphodiester bond, with hydrolysis of ATP to AMP and PPi. Substrates include singlestranded RNA and DNA as well as dinucleoside pyrophosphates (1).

产品来源

An E. coli strain that carries the T4 RNA Ligase 1 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
    • M0437M     -20    
        T4 RNA Ligase 1 (ssRNA Ligase), High Concentration M0437MVIAL -20 1 x 0.167 ml 30,000 units/ml
        T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X
        PEG 8000 B1004SVIAL -20 2 x 1 ml 1 X
        ATP N0437AVIAL -20 1 x 0.1 ml 100 mM

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to convert 1 nanomole of 5´- [32P rA16 into a phosphatase-resistant form in 30 minutes at 37°C

    反应条件

    1X T4 RNA Ligase Reaction Buffer 
    Supplement with 1 mM ATP
    Incubate at 25°C

    1X T4 RNA Ligase Reaction Buffer 
    50 mM Tris-HCl
    10 mM MgCl2
    1 mM DTT
    (pH 7.5 @ 25°C)

    贮存溶液

    50 mM KCl
    10 mM Tris-HCl
    0.1 mM EDTA
    1 mM DTT
    50% Glycerol
    pH 7.5 @ 25°C

    热失活

    65°C for 15 minutes

    单位活性检测条件

    1X T4 RNA Ligase reaction buffer, supplemented with 1 mM ATP, is mixed with the RNA substrate (10μM of 5´-[32P]rA16 ) and varying amounts of enzyme. Incubation is at 37°C for 15 minutes (8).

  • 优势和特性

    应用特性

    • Ligation of ss-RNA and DNA
    • Labeling of 3´-termini of RNA with 5´-[32P] pCp (3)
    • Inter- and intramolecular joining of RNA and DNA molecules (4,5)
    • Synthesis of single-stranded oligodeoxyribonucleotides (6)
    • Incorporation of unnatural amino acids into proteins (7)

  • 相关产品

    相关产品

    • m0204-t4-rna-ligase-1-ssrna-ligase

  • 注意事项

    1. pCp 连接需要加入终浓度 10%（v/v）的 DMSO（3）。

  • 参考文献

    1. England, T., Gumport, R. and Uhlenbeck, O. (1977). Proc. Natl. Acad. Sci. USA. 74, 4839-4842.
    2. Rand, K.N. and Gait, M.J. (1984). EMBO J. 3. 397-402.
    3. England, T. and Uhlenbeck, O. (1978). Nature. 275, 560-562.
    4. Romaniuk, P. and Uhlenbeck, O. (1983). In R. Wu, L. Grossman and K. Moldave(Ed.), Methods in Enzymology. Vol. 100, (pp. 52-56). New York: Academic Press.
    5. Moore, M.J. and Sharp, P.A. (1992). Science. 256, 992-997.
    6. Tessier, D.C., Brousseau, R. and Vernet, T. (1986). Anal. Biochem. 158, 171-178.
    7. Noren, C.J. et al. (1989). Science. 244, 182-188.
    8. Silber, R., Malathi, B.G. and Hurwitz, J. (1972). Proc. Natl. Acad. Sci. USA. 69, 3009-3013.

操作说明、说明书 & 用法

  • 操作说明

    1. Ligation of a DNA or RNA oligo to the 3’ end of long ssRNA using High Conc. T4 RNA Ligase 1 (NEB #M0437)

工具 & 资源

  • 选择指南

    • Substrate-based Ligase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. What is the optimal reaction temperature and time for T4 RNA Ligase I?
    2. Can T4 RNA Ligase 1 be used to end label single stranded DNA or RNA?
    3. Will T4 RNA Ligase 1 ligate double stranded DNA?
    4. What is the most common cause of ligation failure when using T4 RNA Ligase 1?
    5. Why doesn’t the new T4 RNA Ligase reaction buffer contain ATP?
    6. Will T4 RNA Ligase 1 ligate single stranded DNA?
    7. Will the ligation reaction using T4 RNA Ligase 1 produce circles or duplex products?
    8. Can T4 RNA Ligase 1 be used to make single stranded RNA/DNA hybrids?
    9. Can T4 RNA Ligase 1 ligate triphosphates?
    10. Can T4 RNA Ligase 1 be used to add tails to linear duplex DNA?
    11. Can a dideoxy be used to block ligation using T4 RNA Ligase 1?
    12. Can T4 RNA Ligase 1 be used to incorporate unnatural amino acids into proteins?
    13. Will PEG improve ligation with T4 RNA Ligase 1?
    14. Is PEG 8000 available for purchase?