PGLO Bacterial Transformation Lab Assignment If the genetically transformed cells have acquired the ability to live in the presence of the antibiotic ampicillin, then what might be inferred about the other genes on the plasmid that you used in your transformation procedure?
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You transform bacteria with a plasmid encoding a gene that confers resistance to the antibiotic kanamycin. You plate this mixture along with other controls on different agar plates. These are described below: What was plated? Plate 1: Cells Transformed with Water Plate 2: Cells Transformed with Plasmid Plate 3: Water Plate 4: Cells Transformed with Water Plate 5: Cells Transformed with Plasmid Plate 6: Water Kanamycin on plate? Plate 1: Y Plate 2: Y Plate 3: Y Plate 4: N Plate 5: N Plate 6: N Is there growth? Plate 1: N Plate 2: N Plate 3: N Plate 4: Y Plate 5: Y Plate 6: Y Which of the following statements is consistent with these observations? Select one: a. The bacterial cells you are using are not resistant to antibiotic without the plasmid. b. The bacterial cells you are using are not viable. c. There was no bacterial contamination from the environment in your experiment. d. Bacteria were successfully transformed with your plasmid of interest.
Josee P.
Speculate the results you would observe if you inserted a piece of DNA into the Multiple Cloning Site of the pDrive plasmid, then transformed bacteria as we did in this lab. The piece of DNA is inserted into the MCS and then it is transformed into bacteria. Inside the bacteria, the plasmid starts its expression and produces a protein product of our DNA interest. At the same time, we identify which bacterial colonies contain the plasmid by adding X-Gal, a medium with TPTG. The X-gal degrades beta-galactosidase and turns the colonies blue. This shows that the plasmids have been transformed into bacteria.
Adi S.
Briefly explain how an antibiotic-resistance gene and the lacZ gene can be used to determine which cells contain a particular plasmid.
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