lab2 - A sample was immediately taking from the cocktail,...

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Chart 1: PAHBAH Reaction Controls Reaction Cocktails Color Positive Control Dark Yellow Negative Control Clear This test was done to ensure that our PAHBAH assay was working properly. The positive control is a known reducing sugar (glucose) mixed with a p -hydroxybenzoic acid hydrazide (PAHBAH) solution, and the negative is distilled water mixed with a PAHBAH solution. The cocktails were vortexed, then placed into a boiling water bath for 2 minutes. The positive control turned dark yellow, and the negative control remained clear. This proved that the PAHBAH assay was, indeed, working properly. Graph 1: Positive Enzyme Reaction This test was performed to find out if the reagents for the enzyme reaction (the buffer, the substrate, and the enzyme) are properly made an working (i.e., they will effectively catalyze the conversion of sucrose to reducing sugars.) The reaction cocktail was made of 2 ml pH 4.4 buffer, 2 ml sucrase, and 2 ml 1% sucrose solution.
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Unformatted text preview: A sample was immediately taking from the cocktail, resulting in a Time=0 sample. This was used to blank the spectrophotometer before reading the absorbance rates of the other samples. Then, at 2 minute intervals, .5 ml was taken from the reaction cocktail and put into a test tube containing 2 ml of PAHBAH solution (to react with reducing sugars), then incubated in a boiling bath for two minutes. The graph above shows the quantitative results for the products of the sucrase reaction based on different increments of time. Graph 2: Parameter 1 For the Optimum Temperature experiment, we made the following cocktails for each temperature: 2ml buffer, 2ml enzyme (sucrase), 2 ml substrate (sucrose), as well as a negative control consisting of 2 ml buffer, 2 ml distilled H20, and 2 ml substrate (sucrose.) The temperatures that these cocktails were made for are 0...
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lab2 - A sample was immediately taking from the cocktail,...

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