Genetic Transformation : Effects Of Introducing The Gfp Gene Into E. Coli Bacteria
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Life 102 L 17
2 December 2014
Genetic Transformation: Effects of Introducing the GFP Gene into E. Coli Bacteria
When a bacterial cell incorporates foreign DNA into its own genome, this is known as bacterial transformation, a form of genetic transformation. There are several ways to transform genetic material; projectile bombardment, electroporation, and heat shock. This experiment uses the heat shock method. Interest in genetic transformation began as early as 1928 when Frederick Griffith performed an experiment with pneumococcus bacteria and mice, which suggested the possibility of bacterial transformation (Griffith 1928). Expounding upon this discovery, other experiments proved that DNA was…show more content… This experiment demonstrates genetic transformation and the way genes are expressed, which is important for a multitude of reasons. Gene transformation can be used in many ways, including but not limited to: crop modification, medical research, and bioremediation.
Materials and Methods: For this lab we used two microcentrifuge tubes, a pair of vinyl gloves to prevent contamination, a micropipetter, six sterile micropipetter tips, 500 µl of a transformation solution containing CaCl2, seven sterile loops, ice, a timer, a 42°C water bath, a floating rack two colonies of E. Coli bacteria, pGLO plasmid DNA, four Luria Bertani (LB) broth nutrient agar plates (one with LB alone, two with LB and ampicillin, and one with LB, ampicillin, and arabinose), 500 µl of LB nutrient broth, tape, a UV light, and a 37°C incubator. We labeled the two microcentrifuge tubes, one “+pGLO” and the other “-pGLO”, then put on our gloves. We pipetted the 500 µl of transformation solution into these two tubes, half (250 µl) in each, and put these tubes on ice. Then we put a colony of E. Coli bacteria into each tube and put pGLO plasmid DNA into the +pGLO tube, using sterile loops. We put the tubes back on ice, timing it for 10 minutes. During the interim, we retrieved and labeled the bottom of each of the four LB nutrient agar plates, according to what they contained and would contain. We then took both tubes out of the ice and immediately placed them into a floating rack