Bacterial Transformation Lab

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Arizona State University *

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130

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Biology

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Apr 3, 2024

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docx

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Uploaded by MasterLightning2689

Bacterial Transformation Lab The goal of this assignment was to transport genes from one organism to another with the help of plasmid. The procedure used a procedure to transform bacteria with a gene that codes for Green Fluorescent Protein. Our hypothesis was that the -PGLO LB/amp was going to have the most bacterial growth out of the 4 bacteria’s. During this lab, we hope to see which plate is going to have the most bacterial growth. The lab is focusing on genetic transformation, and the literal meaning is a change caused by genes. Scientists use genetic transformation for many areas of biotechnology. One example of this is transforming a sick person’s cells with healthy copies of the defective gene that causes the disease. The hypothesis that will be tested in the lab is if -PGLO LB/amp will have the most bacterial growth or if another will. After we transport the protein plasmid into the bacteria’s, we hope to see clear evidence to which one grew the most bacteria. The materials needed for this assignment includes tubes, pipette, the bacteria’s, a sterile loop, plates, and plasmid DNA. To start, you will need the two test tubes of transformation solution, one +pGlO solution and the other -pGLO. Then using a sterile loop, pick up 2-4 colonies of the bacteria from the starter plate. Put the colonies of bacteria into the +pGLO solution and spin the loop until the colony is dispensed in the transformation solution. Then repeat with the –pGLO tube. After, examine the pGlO solution under the UV lamp. Then using a

sterile pipet, add 250 ul of LB nutrient broth to both tubes. After, pipet 100 ul of the transformation to the 4 agar plates. Once done, you will need to spread the suspensions evenly around the surface by taking a sterile loop and spreading the solution across the plate surface. Finally, stack the plates and let them incubate. Our controls for the experiment were the –pGLO LB/amp and the -pGLO LB plates. The independent variable was the plasmid, and the dependent was the bacteria. Results: Discussion: Once we received our plates back, we were quite surprised. We could see that + LB/amp ara and + LB/amp both had a significant amount of growth of bacteria. On the other hand, - LB and -LB amp had few amounts of bacterial growth on their plates. This was contrary to our

hypothesis as we predicted that -LB amp was going to have the most bacterial growth. Some errors that could have happened in the lab was when spreading the solution over the plates with the loop, we pressed too hard and made a dent in the plates. To avoid this, gently spread the solution over the plates and make sure not to press down on the utensil.

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