Analysis : ' A ' Glowing, Gone ! : Bacteria Transformation And Who Dunnit?

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Austin, Mariya, Kaitlin Curran AP Bio 11 December 2014 Glowing, Glowing, Gone!: Bacteria Transformation and Who Dunnit?: Electrophoresis Analysis with DNA Purposes: · Describe the process of transformation in bacteria · Understand how to prepare viable E. coli cells · Describe the components of gene transfer using plasmid vectors · Be able to describe how antibiotic resistant genes are able to transfer, and identify the transformed cells that are antibiotic resistant · Describe how restriction endonucleases work · Describe how a piece of foreign DNA can become a part of a plasmid that ends up altering the final phenotype of cells · Explain the process of electrophoresis as it relates to separating and identifying different DNA fragments Hypothesis: If a gene that codes for Green Fluorescent Protein transforms bacteria and GFP glows when transformation occurs, then when two micro test tubes have 250 microliters of transformation solution and places in an ice bath, then 2-4 bacteria colonies are added to each tube with a sterile loop; then a plasmid (pGLO) is added to one of the tubes, incubated in ice for 10 minutes, then heat shocked for 50 seconds at 42 degrees Celsius, then back into 9ice for two minutes; then LB nutrient broth is added to both tubes (250 microliters) and set out at room temperature for 10 minutes. Then, 100 microliters of each solution in the tube are added to four

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