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Lab3_09_ - Author Jack Reifert Page 1 Week 3(Oct 12-16...

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Author: Jack Reifert Page: 1 Week 3 (Oct 12-16) Laboratory #3 Quiz #1 this week in lab! Good Luck! Lab #3 introduces two experiments to analyze enzyme activity. The first experiment measures the enzymatic production of a colored product. In the last lab we learned how to use a spectrophotometer to measure absorbance of a colored dye, which we will apply for this experiment. The second experiment in today’s lab utilizes enzymes that cut up DNA. In this lab we run the experiment with these enzymes and save the samples for Lab 4 when a method to visualize DNA is introduced. Finally, the results from this DNA visualization will be analyzed in the lab #5. Part I) Enzyme Activity: - Main Point: Use of a colometric assay to visualize the activity of an enzyme (Tyrosinase) - Enzyme conversion of a substrate into a product - Point: Enzymes as catalysts to speed up reaction rates inside the cells - Left on their own, the reactions inside cells would either never occur or occur too slowly for life. This is why enzymes are needed to help these reactions along. - Enzymes lower the activation energy for the reaction making it easier to get over the “hump” - Structure vs. Function : the structure of the enzyme is necessary for its function of making dopachrome. - Practice using micropipettors - Know which pipetteman for the different volumes - Always use tips on the pipettors Recall absorbance and color wheel parts of lab 4… This is putting that knowledge to use. - Basic Steps: For the extraction section, I will make a big batch of mushroom “slurry” for everyone to use. 1) Make a mixture of mushrooms and phosphate buffer - The buffer is necessary to keep the enzymes we wish to extract at the correct pH so they can function. 2) Blend the mixture to make a “slurry” and extract the enzyme from the mushrooms 3) Filter the “slurry” to obtain a more pure extract
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