The CVA10 strain CVA10-FJ-01 (GenBank accession no. KY012321) had been isolated from a clinical patient in 2014 in Fujian Province and was grown in RD cells (American Type Culture Collection, CCL-136) at a multiplicity of infection of 0.1. Infected cells were incubated at 37°C for 3 days and centrifuged at 7000g for 30 min to remove cell debris. The virus in the supernatant was precipitated using 6% (w/v) PEG 8000 in PBS buffer (pH 7.4). After one more centrifugation, the virus was resuspended in PBS buffer and subsequently loaded onto a 15 to 45% (w/v) sucrose density gradient for ultracentrifugation in a Beckman SW41 Ti rotor at 150,000g for 3.8 hours. The virus bands were individually collected and dialyzed against PBS buffer. Then, the virus was concentrated with cutoff filters. All steps of the purification procedure were carried out at 4°C. The purity of virus preparation was assessed with Coomasie-stained SDS-PAGE gel (NuPAGE 4-12% Bis-Tris Gel, Invitrogen).

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.