标签归档：ligase

RtcB Ligase |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

产品来源

RtcB Ligase is expressed as a His-tagged fusion in E. coli.

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
    • M0458S     -20    
        RtcB Ligase M0458SVIAL -20 1 x 0.025 ml 15 µM
        RtcB Reaction Buffer B0790AVIAL -20 1 x 0.5 ml 10 X
        MnCl2 B1761SVIAL -20 1 x 1 ml 10 X
        GTP N2080AVIAL -20 1 x 0.05 ml 10 mM

  • 特性和用法

    反应条件

    1X RtcB Reaction Buffer
    Supplement with 0.1 mM GTP 和 1 mM MnCl2
    Incubate at 37°C

    1X RtcB Reaction Buffer
    50 mM Tris-HCl
    75 mM KCl
    3 mM MgCl2
    10 mM DTT
    (pH 8.3 @ 25°C)

    贮存溶液

    50 mM NaCl
    10 mM Tris-HCl
    0.1 mM EDTA
    1 mM DTT
    50% Glycerol
    pH 7.4 @ 25°C

  • 优势和特性

    应用特性

    • Ligate ssRNA or ssDNA with a 3´-phosphate or a 2´,3´-cyclic phosphate to the 5´-OH of ssRNA
    • Circularization of ssRNA with compatible ends, as above
    • Detection of 3´-phosphorylated or 5´ -OH RNA using labeled probes

  • 注意事项

    1. 对于困难底物，添加 15% 的 PEG 8000 可以提高连接的产量。

  • 参考文献

    1. Naoko, T., Shuman, S. (2011). J. Biol. Chem. 286, 7727-7731.
    2. Naoko, T. et al. (2011). J. Biol. Chem. 286, 43134-43143.
    3. Chakravarty, A. K. et al (2012). Proc. Natl. Acad. Sci. USA. 109, 6072-6077.

操作说明、说明书 & 用法

  • 操作说明

    1. Typical Protocol for RNA Ligation (NEB #M0458)

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. Does secondary structure affect ligation by RtcB Ligase?
    2. Can RtcB Ligase ligate DNA?
    3. What is the optimum reaction temperature for RtcB Ligase?
    4. Can RtcB Ligase be used in other NEBuffers?
    5. How can I increase ligation efficiency?
    6. Can magnesium, or another divalent cation, be substituted for manganese?
    7. Does PEG improve ligation efficiency?
    8. Can I purchase large amounts of an existing Enzyme for Innovation?
    9. What are Enzymes for Innovation?

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

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

 

产品来源

An E. Coli strain that carries the T4 RNA Ligase I 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
    • M0204S     -20    
        T4 RNA Ligase 1 (ssRNA Ligase) M0204SVIAL -20 1 x 0.1 ml 10,000 units/ml
        T4 RNA Ligase Reaction Buffer B0216SVIAL -20 1 x 1.5 ml 10 X
        Adenosine-5′-Triphosphate (ATP) B0756AVIAL -20 1 x 0.2 ml 10 mM
        PEG 8000 B1004SVIAL -20 1 x 1 ml 1 X
    • M0204L     -20    
        T4 RNA Ligase 1 (ssRNA Ligase) M0204LVIAL -20 1 x 0.5 ml 10,000 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
        Adenosine 5′-Triphosphate (ATP) P0756SVIAL -20 1 x 1 ml 10 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

    10 mM Tris-HCl
    50 mM KCl
    1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    pH 7.4 @ 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).

    Concentration:
    10,000 units/ml

  • 优势和特性

    应用特性

    • 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 oligodeoxyribo-nucleotides (6)
    • Incorporation of unnatural amino acids into proteins (7)
    • Ligation of ss-RNA and DNA

  • 相关产品

    相关产品

    • m0437-t4-rna-ligase-1-ssrna-ligase-high-concentration
    • m0239-t4-rna-ligase-2-dsrna-ligase
    • m0242-t4-rna-ligase-2-truncated
    • m0351-t4-rna-ligase-2-truncated-k227q
    • 小鼠 RNase 抑制剂

    单独销售的组分

    • T4 RNA 连接酶反应缓冲液
    • 5´-三磷酸腺苷（ATP）

  • 注意事项

    1. pCp 的连接需要加入终浓度为 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). R. Wu, L. Grossman and K. Moldave(Ed.), Methods Enzymol.. 100, 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. 3´ End Labeling of RNA using T4 RNA Ligase 1
    2. RNA Circularization using T4 RNA Ligase 1(NEBM#0204)
    3. Protocol Ligation of an oligo to the 3’end of RNA using T4 RNA Ligase 1(M0204)

工具 & 资源

  • 选择指南

    • Properties of DNA and RNA Ligases
    • RNA 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 the ligation reaction using T4 RNA Ligase 1 produce circles or duplex products?
    7. Can T4 RNA Ligase 1 be used to make single stranded RNA/DNA hybrids?
    8. Can T4 RNA Ligase 1 ligate triphosphates?
    9. Can T4 RNA Ligase 1 be used to add tails to linear duplex DNA?
    10. Can a dideoxy be used to block ligation using T4 RNA Ligase 1?
    11. Can T4 RNA Ligase 1 be used to incorporate unnatural amino acids into proteins?
    12. Will PEG improve ligation with T4 RNA Ligase 1?
    13. Will T4 RNA Ligase 1 ligate single stranded DNA?
    14. Is PEG 8000 available for purchase?

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

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

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?

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

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

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?