screening. In this experiment, “Beer- lambert equation that changes in absorbance is equal to extinction coefficient*path length(of cuvette)* concentration in mol. /L.” Hence, the absorbance of P-nitrophenol over time will be determined in this experiment which gives us the absorbance 405nm, and values are used to determine C and then calculate the rate of reaction and the total enzyme unit(U) in the reaction for the given enzyme for a given volume over time. The enzyme activity is the mole that is converted per unit time.Materials & Methodsmaterials1m Tris-HCl pH 8.010mg/Ml Methyl ParathionEnzymeIncubatorCuvetteCuvette spectrophotometerParaffin SquareMethods:a blank was created by the instructor and the spectrophotometer was operated. Also, a master mix in cuvette was created by the instructor. 895ul50mM Tris HCL5uL10mg/ml Methyl parathion100ulEnzyme
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BTECH 3100 Oweh 2Method The enzyme was added and mixed as close as to spectrophotometer read as possible. The top of cuvette Para filmed and gripped the top and bottom of the cuvette with thumb and fore finger andinverted gently 3-5x to mix the reagents. The cuvette top is Para filmed properly to avoid leakage. The OD was measured in spectrophotometer at 405nm absorbance. The mix was incubated at 37 degrees Celsius for 5 min and OD measured again and recorded again at 405nmThe incubation is repeated for another 5 minutes increments until 30 minutes total incubation time: and values recorded as read. The empty enzyme cuvette is discarded into biohazardTimeOD: 4500.020250.4224100.9644151.1579201.2464251.2935301.3316
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