Pglo Transformation Lab Report

1. A bacterial transformation is when a foreign DNA is inserted into the bacteria's original DNA to alter the genome for a certain outcome. This is usually done by using a plasmid to transfer and incorporate the foreign DNA into the original genome. First, bacterial cells are centrifuged to make a pellet. Then they are shocked with a calcium chloride solution that changes the charge on the cell membrane so that the plasmid DNA may be accepted into the cell. This solution must be chilled so that the cell membrane may heal. After incubating the bacterial cells, they are heat shocked to open the pores in the cell membrane to allow the transformation to occur. After being chilled again in order not to melt the agar, the cells are placed in a medium

A plasmid is a circular piece of DNA that contains genes that are not part of the original DNA of the bacteria. However, when a plasmid is inserted into a bacterial cell, its genes are transcribed and translated into proteins that the bacterial cell creates. In our experiment, we used the pGLO plasmid which encodes for a green fluorescent protein. These traits are visualized under a UV light; therefore, transformed bacterial cells will glow green when exposed to a UV light.
3. The hypothesis for this experiment was that transformed bacterial cells would grow on ampicillin plates and glow green when exposed to UV light. The rationale for this hypothesis was because the plasmid would code for ampicillin resistance and a green fluorescent protein, we would have the outcome explained in the hypothesis.
4. Our predictions were that for the standard protocol plates with agar and ampicillin, there would be growth and the colonies would glow under UV light. On the modified protocol on plates with agar and ampicillin where we changed the time of the heat shock, we expected less growth but the colonies would still glow. On the first negative control plate, there should have been no growth on the plate with agar and ampicillin and regular E. coli cells. On the negative control plate with just agar and E. coli cells there should have been growth but the colonies would not