Essay on Pglo Transformation Lab Report - 1486 Words

Transformation Lab Report Introduction Transformation is the transfers of virulence from one cell to another, through the transferring of genetic material. It was originally postulated in 1928 through the works of Federick Griffith, a British microbiologist. Griffith observed that the mutant form, non-virulent form, of the bacteria Streptococcus Pnumoniae could be transformed into the normal, virulent form, when injected into mice along with heat killed normal forms. He concluded that somehow the information the dead virulent form had transformed the mutant form into a virulent form. Later on through the works of Avery, Macleod, and McCarty in 1944, it became obvious that DNA is the transforming property and the substance†¦show more content†¦Spin the two tubes in a centrifuge for 5 minutes on opposite side of the centrifuge. The bacterium will collect at the bottom of the tube, so pour out the extraneous supinate. Then, add 250 microliters of buffer. The Ca2+ cation of the buffer neutralizes the repulsive negative charges of the phosphate backbone of the DNA and the phospholipids of the cell membrane allowing the DNA to pass through the cell wall and enter the cells. Place both tubes on ice. Then add 10 microliters of water into one tube and 10 microliters of plasmid DNA into another tube labeling the one with DNA with a + and the one with water -, and place on ice for 10 minutes. Next heat shocks the tubes for 50 seconds, followed by icing for 10 more minutes. The heat shock increases the permeability of the cell membrane to DNA. Then add 250 milliliters of LB and incubate for 20 minutes. The 20 minute incubation following the addition of LB broth allows the cells to grow and express the ampicillin resistance protein, beta-lactamase, so that the transformed cells survive the subsequent ampicillin selection plates. Plate 100 microliters the + tubes evenly on two plates; 1 of LB and Amp, and one of LB, AMP, and ARA. Plate 100 microliters of the – tubes evenly on two plates; 1 of LB and AMP, and one on LB only. Results Calculations for number of molecules of Plasmid DNA Molecules For initial inflorescence of water and plasmid DNA they did not fluoresceShow MoreRelatedTransformation And Electrophoresis Lab Report1750 Words   |  7 PagesSingee 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. 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