产品信息

• 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
  M0224S     -20       HaeIII Methyltransferase M0224SVIAL -20 1 x 0.05 ml 10,000 units/ml   HaeIII Methyltransferase Reaction Buffer B0224SVIAL -20 1 x 1.5 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 HaeIII restriction endonuclease.

  反应条件

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

  1X HaeIII Methyltransferase Reaction Buffer
  50 mM Tris-HCl
  50 mM NaCl
  10 mM DTT
  (pH 8.5 @ 25°C)

  对照保护实验

  HaeIII Methyltransferase is incubated with 1 µg of λ DNA in 10 µl of 1X HaeIII Methyltransferase Reaction Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection is determined by the addition of 40 µl NEBuffer 2.1 and 10 units of HaeIII restriction endonuclease. Incubation for 1 hour at 37°C is followed by analysis on an agarose gel.

  贮存溶液

  50 mM Tris-HCl
  1 mM DTT
  10 mM EDTA
  200 µg/ml BSA
  50% Glycerol
  50 mM KCl
  pH 7.4 @ 25°C

  热失活

  65°C for 20 minutes

• 相关产品

  单独销售的组分

  • S-腺苷甲硫氨酸（SAM）

• 注意事项

  1. HaeIII 甲基转移酶能保护 DNA 不被 NotI 切割。

• 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. Does HaeIII Methyltransferase require DTT?
  4. Can HaeIII Methyltransferase be heat inactivated?
  5. Does HaeIII Methyltransferase require MgCl₂?
  6. What is the molecular weight of HaeIII Methyltransferase?
  7. Will any NEB methyltransferases (methylases) work on RNA?