0.3% formaldehyde was used to cross-link and glycine solution was used to quench the cells (Millipore). Sangon Biotech synthesized the biotin-labelled circDLC1 probes and control probes (Table S6). Biotin-labeled RNA pull-down was conducted according to the manufacturer's instructions of the EZ-Magna ChIRP RNA Interactome Kit (Millipore). Briefly, 1/5 associated RNA-protein complexes were separated for RNA purification, and the remaining were subjected to protein purification. RNA was purified with RNeasy Mini Kit (Qiagen Inc.). RT-qPCR was used to identify and quantify the enrichment of circDLC1. On the other hand, 4/5 associated RNA-protein complexes were treated with RNase A, RNase H and DNase to elute proteins. The purified proteins were subsequently analyzed by Western blot.

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.