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Learn about the discovery of GFP and the 2008 Nobel Prize in Chemistry award by watching this 9 minute video clip:
pGLO Questions:
- What is the purpose of the starter plate?
- Do E.coli typically glow under the presence of UV light?
- What are the three major genes located in the recombinant plasmid and what is the purpose of each gene?
- What is the purpose of the transformation solution?
- What is the purpose of the heat shock?
- What is the purpose of 10 minute recovery period?
- What is the purpose of each of the two -pGLO plates? What do the results indicate about E.coli’s ability to grow and reproduce in the presence of ampicillin?
- Why was ampicillin added to each of the +pGLO plates?
- Beyond a potential source of energy, what was the purpose of placing arabinose in one of the +pGLO plates?
- Observe the two +pGLO plates. What do the results indicate about each of the colonies on the plates?
- Why are colonies on the +pGLO LB/amp/ara plate glowing?
GFP Questions:
- A colony from each +pGLO plate was transferred to a separate liquid culture tube. Why was a colony chosen from each of the two plates?
- What was the purpose of shaking the liquid culture tubes at room temperature for two days?
- Did one or both tubes glow at the end of 48 hours? Why or why not?
- After observing the liquid culture tubes, you placed the contents of the “+” tube into a microcentrifuge tube and then centrifuged the sample. What was in the pellet? Supernatant?
- What was the purpose of adding lysozyme to the pellet? Why was the sample then placed in the freezer?
- After 24 hours in the freezer, the sample was thawed and again centrifuged. What was in the pellet? Supernatant?
- What is the purpose of column chromatography in this experiment?