Pglo Transformation Lab Report

Genetic transformation is portrayed simply as a ”’change caused by genes’” (DNA) from an external source, which alters the traits within an organism ("pGLO Transformation,"). In this lab, a bacterium called Escherichia coli (E. coli) will be transformed utilizing a gene that codes for Green Fluorescent Protein (GFP) (Urnowey et al., 2017). This particular gene (GFP) comes from the bioluminescent jellyfish, Aequorea victoria, where the protein enables the jellyfish to glow in the dark ( Urnowey et al., 2017). Likewise, if the transformation experiment is done effectively, the bacterial cell, E. coli, will inherit the GFP gene and exhibit the same glow found in the jellyfish (Urnowey et al., 2017). The transformation of the bacterial cells can

The cells that undergo transformation can activate the GFP protein using a sugar called arabinose that is added to the cells' growth medium ("pGLO Transformation,"). Using the aseptic technique, transformed cells will grow on the nutrient agar plates with LB/amp, and appear white on the plates that do not contain the sugar arabinose (Urnowey et al., 2017). If bacteria with +pGLO plasmids are found on the plate with LB/amp/ara, growth will take place and it will glow green under UV light because of the presence of arabinose. Furthermore, +pGLO bacteria that contains the gene for GFP and are resistant to the antibiotic ampicillin, will survive and develop on the plate that has LB/amp. In the control plates, -pGLO bacteria that is susceptible to antibiotic ampicillin will not grow on the plate with LB/amp. On the other hand, the plate containing –pGLO and no amp added (LB), will grow colonies across the surface of the agar

Using an aseptic pipet, 250 µl of transformation solution was added to each micro test tube before setting the tubes on ice. Furthermore, using a sterile loop, one colony was added to each micro test tube and placed back on the ice. Using an aseptic pipet, eight µl of pGLO plasmid was transferred to the +pGLO test tube and both tubes were placed on ice for ten minutes. In the meantime, four agar plates were labelled accordingly: LB/amp (+pGLO), LB/amp/ara (+pGLO), LB/amp (-pGLO), and LB (-pGLO). Using the foam test tube rack as a holder, both the (+) and (-) pGLO test tubes were placed in a water bath set at 42°C for 50 seconds, and then moved quickly back onto ice for two minutes. Using two separate aseptic pipettes, 250 µl of LB broth were added to each micro test tube and mixed gently. Likewise, using two separate, aseptic pipettes for each tube, 100 µl of solution was added to the appropriate agar plate. After, using a new loop for each plate, the solution was spread gently across their surfaces. Lastly, the plates were stacked, taped together, and labelled before placing them upside down in an incubator set at 37°C