Escherichia coli (E. coli) is a normal flora bacterial species found in the lower intestines, which acts as an opportunistic pathogen, especially in hospitals and medical care facilities. The medical industry also battles drug‐resistance in such pathogenic bacteria, which usually occurs with improper use of antibiotic treatment on infections. To evaluate bacterial tolerance to antibiotics over time, this study used E. coli (Strain ATCC 23848) in the presence of Ampicillin, Tetracycline, and Kanamycin Sulfate each of which function by inhibiting cell wall synthesis. Tube dilutions tests were conducted to determine the initial minimum inhibitory concentration (MIC) of each antibiotic required to prevent E. coli cell growth. E. coli present in tubes with antibiotic concentrations less than the MIC were then cultured in non‐lethal doses. Every two weeks an MIC was performed to determine if a change in the MIC had occurred.