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The Transformation of Bacterial DNA

 

            
             Bacteria will cultivate in the presence of pGLO, and the student whose sample contains arabinose will exhibit fluorescence under the UV light even after DNA isolation, quantification, restriction digest, and protein analysis.
             Discussion/Analysis.
             The experiments begin with the transformation of pGLO plasmid in E.coli. We observed growth in each of the three cultures due to the presence of the LB broth and the pGLO. Any bacteria present on the LB/AMP and LB/AMP/ARA plates exhibit successful bacterial transformation, because only the bacteria with the transformed DNA can grow in the presence of ampicillin. The third plate; Figure 3, (LB/AMP/ARA) glows under the UV light due to the presence of arabinose (ARA), it activates the expression of the GFP gene that contributes to fluorescence. .
             For the next lab, the transformed DNA is isolated, and then centrifugation is used to collect a cell pellet to be used in later labs. I had the culture with ampicillin and arabinose, while Mohammed's culture contained no arabinose. Under the UV light, my pellet glowed and his did not. This observation is due to the arabinose in my pellet. .
             The subsequent class involved quantification and restriction digest. After diluting our samples, we measured absorbances at 260 nm and 280 nm. My readings were 0.482 and 0.273 respectively, and the quality of my DNA was 1.76. Mohammed obtained a similar value for his sample (1.79). The DNA concentration for the crude prep is usually not more 1 mg/ml (micro), and any numbers greater than that usually indicates contamination. In our case, the DNA quality reading is expected to be greater than 1 because we have RNA in our samples, which also absorbs at a λ max of 260 nm. The reason why there is RNA in our samples is because we did not use any RNAse or other enzymes to degrade it. .
             The following week involved the use of electrophoresis to separate sample components and isolated the GFP protein.


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