MBIO 3812 Summer 2011 Ex 6-4 Closed-System Growth Purpose: In this exercise growth curves will be generated for Vibrio natriegens grown in a closed system under different environmental conditions and the doubling times of the bacterium under each condition will be determined. NOTE: One side of the lab will incubate their flasks at 37 ° C with shaking (aerated) . The other side of the lab will incubate their flasks at 37 ° C without shaking (stationary) . Because this is a long experiment it will be started before the lab begins. Procedure: 1. One hour prior to the start of the lab 1.5 ml of a Vibrio natriegens culture is added to 2 flasks that each contain 47 ml of brain heart infusion broth (BHI, a very nutrient rich broth) plus 2% NaCl. A sterile sample of BHI + 2% NaCl broth is used to blank the spectrophotometer and then the optical density (OD) of a sample from each of the freshly inoculated V. natriegens cultures is read in the spectrophotometer at a wavelength of 650 nm. This reading is
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