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HpaII |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

大肠杆菌菌株，其携带来自副流感嗜血杆菌（Haemophilus parainfluenzae）（ATCC 49669）的HpaII 基因。

产品类别:
Methylation Sensitive Restriction Enzymes for Epigenetics Products,
Restriction Enzymes for Epigenetic Analysis,
Epigenetic Analysis Products,

Restriction Endonucleases H M Products,

Time-Saver Qualified Restriction Enzymes Products

应用:
Restriction Enzymes for Epigenetics,
5-hmC Detection & Analysis,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,

Restriction Enzyme Digestion

  • 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
    • R0171V     -20    
        HpaII R0171VVIAL -20 1 x 0.1 ml 10,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
    • R0171S     -20    
        HpaII R0171SVIAL -20 1 x 0.2 ml 10,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
    • R0171L     -20    
        HpaII R0171LVIAL -20 1 x 1 ml 10,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
    • R0171M     -20    
        HpaII R0171MVIAL -20 1 x 0.2 ml 50,000 units/ml
        rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

  • 特性和用法

    单位定义

    一个单位是指在 50 µl 的总反应体系中，在 37℃ 下，1 小时内酶切 1 µg λ  DNA 所需的酶量。

    反应条件

    1X rCutSmart™ 缓冲液
    Incubate at 37°C

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

    在不同缓冲液中的活性

    NEBuffer™ r1.1: 100%
    NEBuffer™ r2.1: 50%
    NEBuffer™ r3.1: <10%
    rCutSmart™ Buffer: 100%

    稀释兼容性

    • 稀释液 A

    贮存溶液

    10 mM Tris-HCl
    50 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    200 µg/ml Recombinant Albumin
    50% Glycerol
    pH 7.4 @ 25°C

    热失活

    80°C for 20 minutes

    甲基化敏感性

    dam 甲基化: 不敏感
    dcm 甲基化: 不敏感
    CpG甲基化: 阻断酶切

    同裂酶

    BsiSI
    HapII
    MspI

  • 相关产品

    相关产品

    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）

    单独销售的组分

    • rCutSmart™ 缓冲液

  • 注意事项

    1. HpaII 是 MspI 的同裂酶。
    2. 被盐浓度 > 50 mM 的 KCl 抑制。
    3. CpG 甲基化阻断酶切。

操作说明、说明书 & 用法

  • 操作说明

    1. Optimizing Restriction Endonuclease Reactions
    2. Restriction Digest Protocol
    3. Double Digest Protocol with Standard Restriction Enzymes

  • 使用指南

    • Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
    • Activity of Restriction Enzymes in PCR Buffers
    • Cleavage Close to the End of DNA Fragments
    • Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
    • Double Digests
    • Heat Inactivation
    • NEBuffer Activity/Performance Chart with Restriction Enzymes
    • Optimizing Restriction Endonuclease Reactions
    • Restriction Endonucleases – Survival in a Reaction
    • Restriction Enzyme Diluent Buffer Compatibility
    • Restriction Enzyme Tips
    • Single Letter Codes
    • Site Preferences
    • Star Activity
    • Traditional Cloning Quick Guide

工具 & 资源

  • 选择指南

    • Alphabetized List of Recognition Sequences
    • Compatible Cohesive Ends and Generation of New Restriction Sites
    • Dam-Dcm and CpG Methylation
    • DNA Methylation Table
    • Frequencies of Restriction Sites
    • Isoelectric Points (pI) for Restriction Enzymes
    • Isoschizomers
    • NEB Diluent and Buffer Table
    • Recleavable Filled-in 5′ Overhangs
    • Restriction Enzymes for Epigenetics Selection Chart
    • Time-Saver™ Qualified Enzymes
    • Why Choose Recombinant Enzymes?

  • Web 工具

    • Competitor Cross-Reference Tool
    • DNA Sequences and Maps Tool
    • Double Digest Finder
    • Enzyme Finder
    • NEBcutter™ v3.0
    • NEBioCalculator®
    • REBASE®

FAQs & 问题解决指南

  • FAQs

    1. Is HpaII inhibited by salt?
    2. Is HpaII affected by methylation?
    3. What is the molecular weight of HpaII?
    4. Does spermidine increase HpaII activity?
    5. Does HpaII cleave ssDNA?
    6. What is the activity of HpaII at 25°C?
    7. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    8. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
    9. Are HpaII slow sites found in common vectors?
    10. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

  • 问题解决指南

    • Restriction Enzyme Troubleshooting Guide

HpaII Methyltransferase |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

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?