Bacterial transformation is the transforming of a bacterial cell using plasmids spliced with various types of DNA from other cells (Miyazaki, 201). Instead of a nucleus with chromosomes, bacterial cells have one large circular chromosome in their cell. They also have much smaller rings of DNA throughout their cytosol. These smaller DNA rings are the plasmids which one can manipulate and introduce into cells for transformation to occur. (Miyazaki, 203)
One example of such a manipulated plasmid is the pGLO plasmid. This is the plasmid that we used in this lab. We introduced this plasmid into E. Coli bacteria using the heat shock method. This method involves placing the transformation mixture with the bacteria and the plasmid into ice and then into a hot water bath several times. We also made use of a CaCl2 solution which made the cell walls of the cells even more permeable to the plasmids.
Tags: Transformation Lab Report, Bacterial transformation pglo system lab report, Bacterial transformation
[...] We did in fact find a good number of bacterial colonies on this plate. These colonies did not fluoresce under UV light. The final plate, labeled had a lawn of bacterial colonies on it, which also did not fluoresce under UV light. Note: Our group was not instructed to count the amount of colonies present, and thus this data was not recorded before the plates were thrown out. Discussion In this experiment, we set out to test the transformation of E. [...]
[...] (Miyazaki, 203- 205) The hypothesis was that bacteria on a plate with arabinose would fluoresce if they had been transformed, and that bacteria on a plate with ampicillin would still grow if they were transformed. Specifically, the 4 plates set up were as follows: DNA+(bacteria with plasmid) and ampicillin but no arabinose, DNA+ and ampicillin plus arabinose, DNA-(no plasmid) with ampicillin, and DNA- without ampicillin. We predicted that bacteria would grow on both DNA+ plates and the DNA- plate without the ampicillin. [...]
[...] The DNA-/LB/amp plate was the most puzzling one. This plate exhibited many colonies. Hypothetically, there should not have been any bacterial growth on this plate, since it contained non transformed bacteria and ampicillin. The ampicillin should have killed off these bacteria. The best explanation for this that we could think of is defective ampcillin. We heard throughout the lab that the ampicillin was found to be either old or defective in some way, possibly not working correctly to destroy bacterial cells. [...]
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