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LOYOLA COLLEGE (AUTONOMOUS), CHENNAI – 600 034 M.Sc. DEGREE EXAMINATION – MEDICAL LAB TECHNOLOGY SECOND SEMESTER – APRIL 2006 IB 35 BT 2900 - SEPARATION TECHNIQUES Date & Time : 26-04-2006/9.00-12.00 Dept. No. PART A Answer all the questions Max. : 100 Marks (10 2 = 20) 1. What is the fundamental principle and types in chromatography? 2. Which separation technique makes use of the density differences in macromolecules? 3. What is meant by ‘rpm’ in centrifugation? 4. What are the factors that affect sedimentation of particles in a centrifugal field? 5. In paper chromatography, which reagent is used for detection of amino acids and why? 6. What is meant by affinity chromatography? 7. What is the relationship between pore size, percentage of the gel and molecular size of macromolecules in gel electrophoresis? 8. What are the roles of TEMED and APS in PAGE? 9. What are the applications of Southern and Western blotting? 10. Name the media and solutions used for protoplast isolation. PART B Answer any 4 of the following, each within 200 words (4 10 = 40) 11. Suggest a methodology for the separation of a mixture of lipids, proteins and carbohydrates from a complex mixture of macromolecules 12. Explain the basic principles in centrifugation. Mention the types of centrifugation and their working principles. 13. Write short notes on: (a) Zonal Centrifugation (b) Stationary and mobile phases in chromatography 14. Discuss the principle and procedure for isoelectric focusing. 15. Give an account of the paper chromatography technique. 16. Write a note on isolation of DNA. Mention the principles and reagents used. PART C Answer any 2 of the following (2 20 = 40) 17. Explain: (a) Fundamental Separation techniques and their working principles. (10) (b) Differential centrifugation (10) 18. Discuss in detail the principle, procedure and applications of SDS-PAGE. 19. Explain the principle, procedure and applications of PCR. 20. Discuss: (a) Northern Blotting (10) (b) Isolation of plasmids (10) ______________