Pglo Transformation Lab Report

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bstract When new genetic information is incorporated into an organism genome, genetic transformation has taken place, bacterial transformation is a form of genetic transformation and also the easiest to create within a lab due to the single cell nature of the bacteria used. For this particular experiment pGLO plasmid will be incorporated into E. coli bacteria which will give it genes that code GFP and beta-lactamase proteins into the modified genome of the bacteria. For the experiment two plates were mixed with and without the pGLO plasmid; one was the pGLO, LB nutrient broth mixed the ampicillin, and the other was the LB nutrient broth, ampicillin. For a comparison, two plates were made, the first containing the pGLO plasmid mixed with LB…show more content…
The tubes were labeled +pGLO and –pGLO and placed into the foam tube rack. The tubes were then opened and a sterile pipette was used to transfer 250 uL of transformation solution (CaCl2) were put into each tube and placed on ice for 3 minutes. A sterile loop was then used to pick up a single colony of Escherichia coli or E. coli (dependent variable) form the started plate being sure not to press into the agar. +pGLO was then picked up and the loop of bacteria was immersed into the transformation solution at the bottom of the tube. The loop was spun, using your fingers, until the entire colony was dispersed into the transformation solution. The tube was the placed back into the rack on the ice. A new sterile loop was then used, following the same process for the –pGLO tube. Both tubes then sat in the ice for 3 minutes. A new sterile loop was then immersed into the pGLO plasmid DNA (independent variable) stock tube and a loopful was withdrawn. A film of plasmid solution was across the ring. The plasmid DNA was then mixed into the cell suspension of the +pGLO tube only. The tube was then closed and returned to the rack on ice. Plasmid DNA was not added to the –pGLO. The tubes were then incubated for 10 minutes on ice, making sure the tubes made contact with the…show more content…
They were then labeled with the group name, type of plate, as well as weather the E. coli were exposed to the pGLO plasmid or not. Both the +pGLO and –pGLO tubes were then transferred into a water bath set and 42°C for 50 seconds, once again making sure the tubes made contact with the warm water. After 50 seconds both tubes were placed back on ice (being sure that each transfer between ice and water was as quick as possible). The tubes were incubated on ice for another 2 minutes. The tubes were then removed from the ice and placed on the table top. One tube was opened and new sterile pipette was used to add 250 uL of LB nutrient broth that was then mixed and the tube was closed. The same process was then used for the other tube using a new sterile pipette. The tubes then incubated at room temperature for 20
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