Transformation And Electrophoresis Lab Report

1750 WordsJan 16, 20157 Pages
Singee Nguyen Transformation and Electrophoresis Lab Report Purposes Discuss the principles of bacterial transformation. Describe how to prepare competent E. coli cells. Discuss the mechanisms of gene transfer using plasmid vectors. Discuss the transfer of antibiotic resistance genes and tell how to select positively for transformed cells that are antibiotic resistant. Discuss the mechanisms of action for restriction endonucleases. Discuss how a plasmid can be engineered to include a piece of foreign DNA that alters the phenotype of transformed cells. Understand and be able to explain the principles of electrophoresis as they pertain to separating and identifying DNA fragments. Hypothesis If genetic transformation allows the insertion of a gene into an organism to gain the trait that the inserted gene codes for, then when 250 µl of transformation solution(CaCl2) and 2-4 colonies of bacteria, E. coli, are added to two micro test tubes, labelled +pGLO and –pGLO, with a loopful of pGLO DNA solution in the +pGLO tube, and are incubated for 10 minutes, then transferred to a 42ºC heat bath for exactly 50 seconds, and then incubated in ice again for 2 minutes for a heat shock, and then have 250 µl of LB nutrient broth, and lastly 100 µl of the solutions spread across the surfaces of 4 LB nutrient agar plates, LB/amp, LB/amp/ara, LB/amp, and LB, with +pGLO, +pGLO, –pGLO, and –pGLO used, respectively, the plates with the –pGLO solution will not glow under a UV light after they have
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