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Unit 8 Lab Report

Decent Essays

Hypothesis: The purpose of this experiment was to investigate the question will different food sources affect the level of activity of detoxification enzymes in bean beetles? The class alternate hypothesis is different food sources will affect the level of activity of the detoxification enzymes in bean beetles. The null hypothesis is the different food sources will not have any effect on the level of activity of the detoxification enzymes in bean beetles. Experimental design: The independent variables in this experiment were the types of beans (bean 1 was mung beans and bean 2 was adzuki beans) and enzymes assays used. The dependent variables were the alpha naphthyl acetate esterase (ANAE) and beta naphthyl acetate esterase (BNAE) enzyme …show more content…

Part I and II were performed twice, each time for a different set of beatles grown on a different bean. In order to get the crude extract, four adult beetles were placed in a microcentrifuge tube with a homogenizing buffer to be rapidly homogenized. More homogenizing buffer was added, the contents were centrifuged (to separate the contents), mixed, and the mixture was transferred to new microcentrifuge tubes, leaving the physical beetle parts in the old tube. The new tubes were again centrifuged, mixed and 500 microliters of the mixture was added to the completed crude extract tubes. In part II, after the enzyme assays were prepared, the four beetle tubes were separated into two groups (two for the α-naphthyl acetate and β-naphthyl acetate). Homogenizing buffer and 240 microliters of one of the assays were added to the crude extract solution. Once the enzyme assays were prepared, all final solutions were centrifuged, mixed and incubated, the Fast Blue B Salt stain (used to attach to the proteins in the mixture) was added to each mixture, mixed and incubated to allow the reactions time to occur. The experimental prediction was if a specific sample size of bean beetles are grown on two different food sources, then activity level of their detoxification enzymes will be

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