Note: You are seeing this message either because your browser has not loaded our stylesheets, or because your browser does not support stylesheets (CSS). Please upgrade to a relatively modern browser to improve your experience. Not sure what to upgrade to? Try Firefox.
The Science Advisory Board
Screen Name: 
  		Password: 
 

Protocols

Welcome to The Science Advisory Board's extensive database of research protocols for the biomedical sciences. Protocols are organized by techniques and are fully searchable by key word(s). Protocols are divided into a brief description of the methods, a step-by-step overview of the procedure, a collection of recipes, the supplies needed, and any helpful tips. Members submitting protocols will be rewarded through the ViewPoints system.
... back to articles list

Quick Virus/Viron Purification Protocol

Posted on Wednesday, October 29, 2003

Description
This is a quick method for purifying virus from cell culture without the added hassle of sucrose gradients and identifying fractions positive for virus. This protocol is primarily designed for protein analysis of the collected virions.

Procedure
Virus containing cell culture media is filtered through a .45 micron filter to remove cells and debris and layered on a 20% sucrose cushion in an Ultracentrafuge tube. Samples are spun at 35,000 rpm for 1-2 hours. Liquid is removed and pellet is resuspended in apropriate buffer (for western analysis SDS page loading buffer). Pipet up and down to resuspend virus. Virus can typically be dissociated by heating at 90C for 15 min.

Recipes
20% sucrose (w/v) is made in PBS and then filter sterilized. Store at 4C for several months. Check periodically for contamination.


Supplies
Ultracentrafuge, with apropriate rotor and tubes etc
ex. Sw41, beckman Ultra-Clear 14X89 mm tubes for a 2ml cushion with 9mls of media

SDS page buffer, see western protocols.

Heat block or boiling water for protein dissociation.

Tips
Be careful not to mix media with cushion

some cellular debris will be in your sample, so good negative controls are needed to ensure specificity of protein to virus

Virus can be resuspended in other buffers to concentrate and then use for future infections.

Submitted by: bdoehle

Active Members
73,280
Join Today

The Science Advisory Board is the world's original professional network of life scientists.

Members of the SAB:

  • Connect with other scientists.

  • Find tips, methodologies and procedures from established researchers.

  • Share insights, stories, jokes and even "gripes" in an open environment.

  • Voice opinions on companies and products used in their work.

  • Earn generous rewards for their opinions.

Practicing life science researchers and medical professionals participate in The Science Advisory Board's studies, forums, news articles and social media channels.
Get Connected
Steering Committee
Staff