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MicBio 3301 Lab Report #1: Developing a Growth Curve Using a Spectrophotometer and 96-Well Plate Procedures

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MicBio 3301 Lab Report #1: Developing a Growth Curve Using a Spectrophotometer and 96-Well Plate Procedures Abstract The first step to understanding a microbe and manipulating it is to understand its pattern of growth ...

MicBio 3301 Lab Report #1: Developing a Growth Curve Using a Spectrophotometer and 96-Well Plate Procedures Abstract The first step to understanding a microbe and manipulating it is to understand its pattern of growth under specified conditions. Escherichia coli is an abundant microbe that serves many purposes – good and bad. Because of its high prevalence, understanding its growth patterns is highly important. This experiment explores those realms by following procedure to generate a standard curve as well as a growth curve. Two methods, a spectrophotometer and a 96-well plate reader, were used to monitor optical density of an inoculated solution over time. To observe and quantify growth (viable count) over time, dilutions of the inoculum were plated and incubated and the colonies were late counted to determine colony forming units per milliliter (CFUs/ml). These results produced a standard curve as well as the two components of a growth curve. The results revealed that as the incubation time increased, the cell concentration within the suspended solution increased which therefore increased the optical density value. The growth curve showed evidence of the lag phase as well as the exponential phase. The curves also showed less definition in the 96-well plate results compared to the spectrophotometer results due to the small volumes used. However, the 96-well plate method produced more useful data overall. References (1) Doyle MP, Schoeni JL. Survival and Growth Characteristics of Escherichia coli Associated with Hemorrhagic Colitis. Applied and Environmental Microbiology. Oct. 1984; 48(4): p.855-856. (2) Smith I. Mycobacterium tuberculosis Pathogenesis and Molecular Determinants of Virulence. Clinical Microbiology Reviews. July 2003; 16(3): p.463-496. (3) Smith S, Schaffner DW. Evaluation of a Clostridium perfringens Predictive Model, Developed under Isothermal Conditions in Broth, To Predict Growth in Ground Beef during Cooling. Applied and Environmental Microbiology. May 2004; 70(5): p.2728-2733.

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