pGLO & GFP

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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:

  1. What is the purpose of the starter plate? 
  2. Do E.coli typically glow under the presence of UV light?
  3. What are the three major genes located in the recombinant plasmid and what is the purpose of each gene?
  4. What is the purpose of the transformation solution?
  5. What is the purpose of the heat shock?
  6. What is the purpose of 10 minute recovery period?
  7. 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?   
  8. Why was ampicillin added to each of the +pGLO plates?
  9. Beyond a potential source of energy, what was the purpose of placing arabinose in one of the +pGLO plates?
  10. Observe the two +pGLO plates.  What do the results indicate about each of the colonies on the plates?   
  11. Why are colonies on the +pGLO LB/amp/ara plate glowing? 

GFP Questions:

  1. 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?                                                      
  2. What was the purpose of shaking the liquid culture tubes at room temperature for two days? 
  3. Did one or both tubes glow at the end of 48 hours?  Why or why not?
  4. 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?
  5. What was the purpose of adding lysozyme to the pellet?  Why was the sample then placed in the freezer?
  6. After 24 hours in the freezer, the sample was thawed and again centrifuged.  What was in the pellet?  Supernatant?
  7. What is the purpose of column chromatography in this experiment? 

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