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Formal Lab Report 2

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Sean Cui
Biology
Formal Lab Report 2

Intro Throughout this lab, the main topic is genetic transformation. Genetic Transformation is a phenotypic change caused by genotype change from transferring foreign genes into an organism. This lab is to transform the bacteria E.Coli with the GFP gene (Green Fluorescent Protein). GFP makes organisms glow in the dark with UV light, similar to jellyfishes.GFP will be transferred into E.Coli through the use of recombinant DNA, or simply a vector. A vector is a plasmid that transfers foreign DNA into cells. Bacteria can transfer plasmid to other bacteria so they can survive and adapt to the environment they are in. The pGLO plasmid includes the GFP gene (allows bacteria to glow), Beta-lactamase (resists ampicillin), ORI sites (allows bacteria to self replicate), and AraC (allows induction of GFP gene).
Materials and Methods: In this lab, first there will be two micro test tubes, one labled pGLO+ and the other pGLO-. Using a sterile transfer pipet, transfer 205 microliters of CaCl2 into each tube. Then put the tubes on ice. Then use a sterile loop to pick up a single colony of bacteria from the starter plate. Put the colony in the pGLO+ tube and spin the loop. Then put the tube back in ice. Repeat this for the pGLO- tube with a new sterile loop. Next use a new sterile loop and get some DNA stock. With the loop, immerse it in only in the pGLO+ tube. After that, incubate the tubes in ice for 10 minutes. Label one plate LB/amp +pGLO, another LB/amp/ara +pGLO, another LB/amp –pGLO, and the last one LB-pGLO. Heat shock the test tubes in 42 degrees celcius for 50 seconds and incubate them in the ice bath for 2 more minutes. After that, put 250 microliters of LB broth into each tube and incubate in room temperature for 10 minutes. Then transfer 100 microliters of the content in each tube to their appropriate plates. Stack

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