Download Activity 4.1.3 Student Response Sheet

Activity 4.1.3 Student Response Sheet
Answer the questions below as you work your way through each step of the GFP
Purification laboratory. Be sure to think about how all of the steps function together to
isolate the protein of interest.
NOTE: The experiment begins with Lesson 2. Lesson 1 covers the background
information for the lab experiment and not the actual hands-on part of the work.
Lesson 2 – Inoculation
Goal:
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1. What would happen if you took one of the green colonies from your LB/amp/ara
plate and streaked it on a LB/amp plate? Conversely, what would happen if you
took a white colony from the LB/amp plate and streaked it on an LB/amp/ara
plate? Explain your answer.
2. Explain how an incubator helps with this cloning experiment.
Lesson 3 – Purification Phase 1: Bacterial Concentration
Goal:
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1. Explain the role of lysozyme and the freezing of the cells play in the isolation the
GFP from the other bacterial proteins.
© 2010 Project Lead The Way, Inc.
Medical Interventions Activity 4.1.3 Student Response Sheet – Page 1
2. Explain why both liquid culture tubes fluoresce green.
3. Why did you discard the supernatant and keep the pellet in this part of the
experiment?
Lesson 4 – Purification Phase 2: Bacterial Lysis
Goal:
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1. What color was the pellet in this step of the experiment? What color was the
supernatant? What does this tell you?
2. Why did you discard the pellet and keep the supernatant in this step of the
protein purification procedure?
Lesson 5 – Purification Phase 3: Protein Chromatography
Goal:
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© 2010 Project Lead The Way, Inc.
Medical Interventions Activity 4.1.3 Student Response Sheet – Page 2
1. Which buffer, the binding buffer, the wash buffer, or the elution buffer has the
greatest salt concentration? Explain your answer.
2. Explain the role or function of each buffer listed in the previous question. How do
they work together to help you isolate GFP?
3. Were you successful in purifying and isolating the desired compound from the
bacterial proteins? Provide evidence to support your answer.
© 2010 Project Lead The Way, Inc.
Medical Interventions Activity 4.1.3 Student Response Sheet – Page 3