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.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.