Viral Plaque Assay Protocol

This template was adapted from the original submission. Edits were made to enhance scientific accuracy, optimal usability and/or to meet industry-leading design standards for science communication. This figure describes a general protocol for performing a viral plaque assay. First, cells are grown to near confluency in a 12 well plate. Once cells reach confluency, media is removed and 50 μL of serially diluted virus is added to each well. Phosphate-buffered saline (PBS) is added to one well as a control. Following 1 hr incubation at 37°C and 5% CO2, 3 ml of growth media overlay agar is added to each well, and the wells are incubated for 48 hrs at 37°C and 5% CO2. Afterwards, 1 ml of 4% PFA is deposited onto the overlay and left for 1 hr at room temperature. Lastly, 1 ml of crystal violet is added to each well and incubated for 5 minutes. Wells can then be rinsed and observed.

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