Using dA methylase (DAM) (New England Biolabs, M0222), a methyl group was added to the adenine (N6) in every 5′-GATC-3′ sequence. One microgram of DNA, 1× methyltransferase reaction buffer, 80 μM S-adenosylmethionine, 1 μl of DAM, and distilled water in 10 μl were incubated at 37°C for 1 hour, followed by 65°C for 15 min. The reaction was scaled up as needed. To control for successful methylation, 500 ng of DNA were digested with either Dpn I (New England Biolabs, R0176) or Mbo I (New England Biolabs, R0147) at 37°C for 15 min. Dpn I cuts at 5′-GATC-3′ if 6mdA is present, while Mbo I cuts the same sequence in the absence of adenine methylation. The resulting products were run on a 1% agarose gel.

注意:以上内容是从某篇研究文章中自动提取的,可能无法正确显示。



Q&A
请登录并在线提交您的问题
您的问题将发布在Bio-101网站上。我们会将您的问题发送给本研究方案的作者和具有相关研究经验的Bio-protocol成员。我们将通过您的Bio-protocol帐户绑定邮箱进行消息通知。