Introduction Genetic transformation involves E.coli cells being injected with new DNA, allowing new traits to be developed. ("pGLO Bacterial Transformation Kit | Life Science Education | Bio-Rad," n.d.) Plasmids, known as bacteria that contains one or more small pieces of DNA, allows more than just one trait obtained from plasmid DNA genes. The process known as genetic engineering is used to insert certain genes that will code for new traits into the plasmid. The pGLO plasmid was manufactured with a jellyfish gene that encoded the green fluorescent protein, an antibiotic-resistance gene that encodes β-lactamase protein, and the araC gene encoding a regulator protein that will turn the GFP gene on or off. In this experiment, pGLO plasmid was …show more content…
Both were then placed into a foam tube rack. After being placed on a foam tube rack the tubes were opened and a sterile pipet was used to transfer 250 microliters of transformation solution (CaC1₂) into both tubes. These tubes were then placed on crushed ice. A sterile loop was used to pick up a single colony of bacteria from the starter plate. The +pGLO tube was then picked up and the loop was immersed into the transformation solution at the bottom of the tube. The loop was spinned between the index finger and the thumb until the entire colony was dispersed in the transformation solution. The tubes were placed back into the rack of ice. Using a new sterile loop, the same process was repeated for the -pGLO tube. The pGLO plasmid DNA solution was then examined under a UV lamp. A new sterile loop was immersed into the plasmid DNA stock tube. When the loop was withdrawn, a film of plasmid solution appeared across the ring of the loop. Afterwards the loopful was mixed into the cell suspension of the +pGLO tube. A pipet of ten microliters of pGLO plasmid was placed into the +pGLO tube and mixed. The -pGLO tube was closed and put back into the the rack on ice. Since plasmid was not added to the -pGLO tube it was closed and returned back into the rack on ice. The tubes were incubated on ice for about ten minutes. While the tubes sat on ice, four agar plates were labeled. The