Take a previously prepared microplate with anti-fog solution and agarose. Pipette 75 microliters of the bacteria into columns three to 10. Next, add 75 microliters of sterile phage buffer to wells in columns three and four as a no-phage positive control.
Mix the solution by pipetting up and down after each addition. Then add 75 microliters of the phage at varied concentrations to the columns, and mix each solution by pipetting up and down again after each addition. Stick both short sides of the plate using 0.5 inch labeling tape, which will partially seal the plate while allowing for gas exchange.
Incubate the plate on a microplate shaker, and measure optical density using a microplate reader. Then, with the optical density measurements, generate control and infected growth curves. The growth curve depicts a productive phage infection shown by the reduced bacterial absorbance over time in wells with the phage added.
This reduction in bacterial density will not be seen if the bacterium is outside the phage's host range.
