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Quantification of a Diet Coke: Standard Ames Test

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Quantification of a Diet Coke: Standard Ames Test
LAB #11A: Quantification of a Mutagen – Standard Ames Test
Sarah R. Villarreal
Rogers 1130

INTRODUCTION.

In the experiment Lab 6A, Diet Coke was determined a mutagen by using a test called the Spot-Overlay Ames Test. The Spot-Overlay Test was designed and named after Dr. Bruce Ames, a Scientist and professor of Biochemistry and Molecular Biology at an array of colleges (Ames 2012), the test was designed to determine whether or not something is mutagenic. After the Spot-Overlay Ames Test we then wanted to know at what concentration Diet Coke became mutagenic so we used the Quantitative Ames test. The quantitative Ames test is similar to the spot-overlay Ames test, except that the mutant Salmonella tymphimuium (which cannot composite histidine) is treated with various concentrations of the mutagen Diet Coke to view which sections of the Davis Minimal Agar Plate form colonies (Bjeldanes, et al. 1982). Mutagenicity has been known to be carcinogenic and that can cause a toxicological effect that presents a potential health risk for humans ( Resende, et al. 2012). The purpose for doing this experiment was to see at exactly what point Diet Coke becomes mutagenic.
Null hypothesis: Diet Coke will not be mutagenic at all concentrations.
Alternative hypothesis: Diet coke will be mutagenic at all concentrations.

MATERIALS & METHODS

We used four different concentrations of the mutagen Diet Coke to test at exactly what percentage Salmonella tymphimurium becomes mutagenic. The treatments were 100% concentrated Diet Coke, 50% concentrated Diet Coke, 25% concentration Diet Coke, and 5% concentration Diet Coke. The plate used for the experiment was the Davis Minimal Agar plate which is a soft gel-like plate that was divided into four sections for each of the treatments to reduce the risk of cross-contamination. There were several controls in this experiment: the positive control and the negative control, as well as the experiment control and the



Cited: 1. Ames, Bruce N. "Nutritional Science & Therapy." J Nutr 2.7-72 (2012). 2. Bjeldanes, L. F., et al. "Mutagens from the cooking of food. II. Survey by Ames/Salmonella test of mutagen formation in the major protein-rich foods of the American diet." Food and Chemical Toxicology 20.4 (1982): 357-363. 3. Resende, F.A., Vilegas, W., Santos 3, L.C., Varanda, E.A., “Mutagenicity of Flavonoids Assayed by Bacterial Reverse- Mutation (Ames) Test.” Molecules 1420-3049 (2012): 5256-5257.

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