For radiolabeling experiments, HeLa cells were lysed in TENTG-150 buffer, and DNA was extracted with phenol/chloroform/isoamyl (pH 8) (12/12/1). DNA was subsequently dephosphorylated using Shrimp Alkaline Phosphatase (rSAP) New England Biolabs (NEB) before labeling with γ32P ATP for 30 min at 37°C using the T4 polynucleotide kinase (NEB). Subsequent RNaseH treatment was performed with 10, 20, or 40 U following the manufacturer’s protocol. Unbound radiolabeled nucleotides were removed using Illustra Microspin G-50 Columns before resolution on 5% acrylamide, 0.5% tris borate ethylamide gel, and autoradiography. Images were acquired using Thyphoon FLA 7000.

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。