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apo223FormalReport - Effectiveness and Efficiency of E coli...

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Effectiveness and Efficiency of E. coli Transformations Under Various Conditions Arria P. Owlia BIO 206, Spring, 2008 March 25, 2008
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Effectiveness and Efficiency of E. coli Transformations Under Various Conditions E. coli can be “transformed” by surrounding plasmids, which, in essence, is the changing of the bacteria’s collections of genes. Once this is accomplished, the bacteria exhibit various traits, for example, our bacteria were to express ampicillin resistance, bioluminescence, and melanin production, although we did not observe bioluminescence in any of our bacteria. Transformation involves the bacteria taking in foreign DNA, plasmids, and ultimately expressing these genes. Upon expression of these genes, one should be able to see the traits that have been added by the plasmids. In our lab, we used three plasmids, Plasmid #1 (pIC 20R), Plasmid #2 (pVIB), and Plasmid #3 (pLAF413). To induce transformation in these bacteria, we dropped the bacteria control and plasmid solutions into a heat bath. We then placed each of the four (Control, Plasmid #1, Plasmid #2, Plasmid #3) solutions onto two agar plates, one without ampicillin, and one with ampicillin. Each plasmid has a gene for ampicillin resistance, thus if there is growth on the agar plate, the bacteria should be transformed. If there is growth on the agar plate without ampicillin, we cannot be sure whether the bacteria have been transformed or not. Our bacteria were incubated after being placed on the agar plates at 37°C for an extended period. After this incubation period, we went ahead and observed the eight agar plates. We checked for three signs: growth, bioluminescence, and color. Our results for the Control group and the Plasmid #1 group were not as we expected. Probably due to human error, the Control and Plasmid #1 groups did not have any growth. For our Plasmid #2 group, we did not observe any bioluminescence, which may be due to the non-transformed bacteria blocking this property to our eyes. Our Plasmid #3 group grew as we had anticipated with melanin production in both, but with a highly concentrated amount of melanin in the agar plate containing ampicillin, because only the bacteria that were transformed were being expressed here. Overall, we learned a few things, including the effectiveness of the transformation-inducing method and the poor accuracy of the human eye.
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Introduction: The purpose of this lab is to gain knowledge and experience in the field of genetics through the use of plasmids and observations of their ultimate effect. Transformation is a widely used technique to obtain improvements in yields with the help of certain genes being introduced to E. coli (Davis et al., 2008). The control of this experiment will be E. coli (DH5 alpha strain) that has not been transformed. The experimental groups involved bacteria that has, hopefully, been transformed by plasmids. There are three plasmids involved in this lab: 1. pIC 20R – plasmid with a gene for ampicillin resistance, 2. pVIB –
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apo223FormalReport - Effectiveness and Efficiency of E coli...

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