HpaII Methyltransferase |NEB酶试剂 New England Biolabs

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产品信息

产品来源

An E. coli strain that carries the cloned HpaII modification gene from Haemophilus parainfluenzae (ATCC 49669).

产品类别:
Methyltransferases for Epigenetics Products,
Methyltransferases Products

  • 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
    • M0214S     -20    
        HpaII Methyltransferase M0214SVIAL -20 1 x 0.025 ml 4,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
        S-adenosylmethionine (SAM) B9003SVIAL -20 1 x 0.1 ml 32 mM

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to protect 1 μg λ DNA in 1 hour at 37°C in a total reaction volume of 10 μl against cleavage by HpaII restriction endonuclease.

    反应条件

    1X rCutSmart™ Buffer
    Supplement with 80 µM S-腺苷甲硫氨酸（SAM）
    Incubate at 37°C

    1X rCutSmart™ Buffer
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 µg/ml Recombinant Albumin
    (pH 7.9 @ 25°C)

    对照保护实验

    HpaII Methyltransferase is incubated with 1 µg of λ DNA in 10 µl 1X rCutSmart Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection by HpaII Methyltransferase is determined by the addition of 40 µl NEBuffer 1 supplemented with 10 mM MgCl2 and 10 units of HpaII restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on agarose gels.

    贮存溶液

    50 mM Tris-HCl
    150 mM NaCl
    0.1 mM EDTA
    5 mM TCEP
    200 µg/ml BSA
    50% Glycerol
    pH 7.5 @ 25°C

    热失活

    65°C for 20 minutes

    分子量

    理论上的: 40397 daltons

  • 相关产品

    单独销售的组分

    • S-腺苷甲硫氨酸（SAM）

  • 参考文献

    1. Hoffman, J.L. (1986). Biochemistry. 25, 4444-4449.

操作说明、说明书 & 用法

  • 操作说明

    1. Recommended Protocol for 3H labeling of DNA
    2. Recommended Protocol for Methylation of Genomic DNA

FAQs & 问题解决指南

  • FAQs

    1. Is S-adenosylmethionine (SAM) supplied with the Methyltransferase?
    2. What should be considered if the methylation is not going to completion?
    3. Can HpaII Methyltransferase be heat inactivated?
    4. Does HpaII Methyltransferase require MgCl2?
    5. What is the molecular weight of HpaII Methyltransferase?
    6. Can HpaII methylated DNA be used to transform E. coli?
    7. Will any NEB methyltransferases (methylases) work on RNA?
    8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?