Precise and High-Throughput Analysis of Bacterial Growth Using a Microplate Reader

Begin with bacteria preserved in a glycerol stock, previously grown to the early exponential phase to ensure uniform physiological states and reproducible growth.

Add growth media and vortex to evenly resuspend the bacteria.

Serially dilute the suspension using fresh media to obtain a range of starting cell densities.

Load the samples into the inner wells of a multi-well plate, leaving the edge wells filled with blank media to minimize evaporation-related variability.

Place the plate in a microplate reader set to maintain constant shaking and a temperature optimal for bacterial growth.

As the bacteria multiply, the optical density or OD of the culture increases.

Measure the OD at fixed intervals.

Subtract the initial OD values obtained from the blank wells to correct for background absorbance.

Then, calculate the growth rate from consecutive OD values measured at defined time intervals, enabling high-throughput and precise quantification of bacterial growth dynamics.

For real time recording of growth, add approximately 25 milliliters of M63 to a sterilized reagent reservoir. Then, add 900 microliters of M63 to the microtubes in preparation for making serial dilutions. Next, add 900 microlit