GFP Column Chromatography
1. In your own words, describe recombinant gene cloning.
2. How were these items helpful in this cloning experiment?
a. UV light
b. Incubator
c. centrifuge
d. lysozyme
e. freezer
3. Explain how placing cloned cells in nutrient broth to multiply relates to your overall goal
of purifying the fluorescent proteins.
4. Explain why the bacterial cell outer membrane ruptures when frozen (what happens to an
unopened soft drink when it freezes)?
5. What was the purpose of lysing the bacteria?
6. Briefly describe hydrophobic interaction chromatography and identify its purpose in this
lab.
7. using your data table, discuss your experimental results. Which fraction contained your
GFP protein?
8. Based on your results, explain the functions of each buffer:
a. equilibration buffer
b. binding buffer
c. wash buffer
d. TE (elution) buffer
GFP Column Chromatography
1. In your own words, describe recombinant gene cloning.
2. How were these items helpful in this cloning experiment?
a. UV light
b. Incubator
c. centrifuge
d. lysozyme
e. freezer
3. Explain how placing cloned cells in nutrient broth to multiply relates to your overall goal
of purifying the fluorescent proteins.
4. Explain why the bacterial cell outer membrane ruptures when frozen (what happens to an
unopened soft drink when it freezes)?
5. What was the purpose of lysing the bacteria?
6. Briefly describe hydrophobic interaction chromatography and identify its purpose in this
lab.
7. using your data table, discuss your experimental results. Which fraction contained your
GFP protein?
8. Based on your results, explain the functions of each buffer:
a. equilibration buffer
b. binding buffer
c. wash buffer
d. TE (elution) buffer
