JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:30 min • July 31st, 2026
Begin with culture supernatants containing rotavirus particles, collected from infected cells at defined time intervals.
Add trypsin to cleave the viral outer capsid proteins, exposing host attachment sites.
Incubate host cells with diluted virus suspensions, shaking intermittently.
The viruses attach to the host and become internalized.
Remove the inoculum and add an agarose medium containing trypsin that supports reinfection.
The agarose solidifies, restricting viral diffusion and ensuring localized infection.
The internalized viruses replicate, leading to cell lysis and release of virions that infect nearby cells.
The virus replication causes cell lysis, forming clear zones or plaques.
Add a dye that stains the living cells, leaving the plaques unstained.
Remove the excess dye and incubate to enhance contrast.
Count the plaques and apply a mathematical model to generate the rotavirus growth curve.
The curve shows a lag phase for adaptation, an exponential growth phase, and a stationary phase when replication slows due to resource limitations.
Store the supernatant at minus 80 degrees Celsius until applying it to the plaque assay for measuring the virus titer.
When ready to perform the plaque
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