Saturday, December 27, 2008
Plaque Assay
Viruses, like other microorganisms are too small to be enumerated by direct counting. Plaque assay can be used to help in counting the viruses.
The theory of the plaque assay is very similar to the theory behind bacterial colony counting. This method requires that the virus infects a cell line that grows as a monolayer. 1 virus will infect one cell if the concentration of virus is low enough. The viral concentration can be determined by the following formula:
Virus concentration = (number of plaques) x (dilution factor(s)) pfu/ml
Plaque Assay: method
Typical Steps to conduct a Plaque Assay
Materials
- 45ml of viral diluent
- 1 vial of 0.5ml kunjin virus suspension
- Sterile test tubes
- Sterile pipette
- 2 6-well trays of confluent BHK-21 cell
- 96ml of viral diluent
- 60ml of 2times concentration maintaince medium
- 60ml of 2% CMC aquacide II Solution in ultrapure water
- Beaker for discard
Procedures
Blogged @ 8:20 PM