Results and Discussion DNA

Results and Discussion DNA - Results and Discussion...

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Results and Discussion Isolating DNA from the Vibrio fischeri bacteria is a comprehensive process involving many reagents. Specific reagents are required to disrupt the various components of the cell wall consisting of an outer membrane containing lipopolysaccharides, a periplasmic space with a peptidoglycan layer, and an inner cytoplasmic membrane. There are many occasions for error to occur if the proper procedures are not followed, even after isolating the DNA malfunction can still occur due to the sensitivity of the DNA. Such a malfunction occurred during this DNA purification process. Spectrophotometric analysis of the isolated DNA solution indicated that the proper amount of DNA was not present. Spectrophotometric analysis is required to estimate the concentration of isolated Vibrio DNA in the solution. This is important to know because calculations could not be done without first knowing the amount of DNA present which will dictate the amount of other reagents to add in later. Concentrations levels will vary every time the purification process is done resulting in a broad range of acceptable results. Estimations of DNA purity from the sample place it in the 1.45 range. Anything below 1.6 or over 2.0 indicates a faulted experiment. Unable to continue with further experiments, group 16 was kind enough to loan their Vibrio DNA samples so that the experiment may continue. Spectrophotometric analysis of group 16’s Vibrio DNA sample gave readings of .067 for OD260 emission and .038 for OD280 emission. Using these results, we were able to determine the concentration, yield, and purity of the sample. Calculation and results are including is the table below. DNA Concentration (ng/microL) Yield Purity Vibrio fischeri 335 335000 .067/.038
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Since now the concentration of the Vibrio DNA sample is known, preparation for Sal I restriction digestion of V. fischeri and pGEM DNA can begin. Since we know the concentration of DNA present we were able to add the correct amount of solution containing the proper amount of DNA. We prepare these solutions in four different tubes. By using the concentration
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Results and Discussion DNA - Results and Discussion...

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