1-What are the advantages and disadvantages of using molecular cloning techniques for purification of proteins? If the desired function is to study in vivo function of a protein, what protein expression system would you use and why?
2-What are the optimization conditions you will have to establish before the purification of a protein and why these optimizations necessary?
3-What are some of the protein monitoring techniques used during protein purification? What is the principle behind each of the techniques you have listed?
4-What is the principle behind molecular permeation chromatography? Give salient features of gel permeation chromatography
5-You have been given a mixture of hemoglobin (Mr. 72 kDa) and cytochrome c (Mr 13.5 kDa), and Sepadex beads (G10, G25 and G75). You have separate and purify these proteins by gel permeation chromatography. Suggest an experimental protocol as to how you would purify them?