|
Lecturer(s)
|
-
Ingr Marek, doc. RNDr. Ph.D.
-
Vícha Robert, doc. Mgr. Ph.D.
|
|
Course content
|
1. Introduction to separation methods, principles of separation. 2. Intermolecular interactions and properties of compounds stemming from them. 3. Processing of complex biological material, basic separation methods (filtration, extraction, precipitation, salting). 4. Electromigration separation methods - principle of zone electrophoresis and isotachophoresis. 5. Survey of capillary and gel electrophoresis methods and their use. DNA sequencing. 6. Sedimentation methods as a tool of analysis of natural compounds - sedimentation velocity in a gravity field and ultracentrifuge. 7. Sedimentation equilibrium, dissociation equilibrium of proteins, isopycnic centrifugation. 8. Chromatographic methods - basic chromatographic principles, analysis of results. 9. Isocratic chromatography methods, gel permeation chromatography - principles and examples of use. 10. Gradient chromatographic methods - ion exchange chromatography, hydrophobic chromatography, reverse phase chromatography, affinity chromatography - principles and examples of use. 11. Principles of detection in chromatographic and electromigration methods - basics of spectrophotometry, fluorometry, refractometry. 12. Detection methods in gas chromatography, mass spectrometry. 13. Methods of natural-compounds analysis - concentration assays, amino acid analysis, protein sequencing. 14. Blotting, PCR and qPCR, immunological methods, flow cytometry.
|
|
Learning activities and teaching methods
|
Lecturing, Simple experiments
- Preparation for examination
- 120 hours per semester
|
| prerequisite |
|---|
| Knowledge |
|---|
| Knowledge of physical and analytical chemistry at the level of bachelor study program, basics of organic and inorganic chemistry, biochemistry, physics and mathematics. |
| Knowledge of physical and analytical chemistry at the level of bachelor study program, basics of organic and inorganic chemistry, biochemistry, physics and mathematics. |
| Knowledge of the basic principles of work in laboratories. |
| Knowledge of the basic principles of work in laboratories. |
| learning outcomes |
|---|
| describe the basic separation methods |
| describe the basic separation methods |
| explain electromigration separation methods |
| explain electromigration separation methods |
| characterise sedimentation methods |
| characterise sedimentation methods |
| sort chromatographic methods |
| sort chromatographic methods |
| define immunological methods |
| define immunological methods |
| Skills |
|---|
| apply electrophoresis for sample analysis |
| apply electrophoresis for sample analysis |
| evaluate the chromatogram |
| evaluate the chromatogram |
| determine the DNA concentration by PCR |
| determine the DNA concentration by PCR |
| apply gel chromatography |
| apply gel chromatography |
| calculate the sedimentation rate |
| calculate the sedimentation rate |
| teaching methods |
|---|
| Knowledge |
|---|
| Lecturing |
| Lecturing |
| Dialogic (Discussion, conversation, brainstorming) |
| Dialogic (Discussion, conversation, brainstorming) |
| Skills |
|---|
| Simple experiments |
| Simple experiments |
| Practice exercises |
| Practice exercises |
| assessment methods |
|---|
| Knowledge |
|---|
| Essay |
| Grade (Using a grade system) |
| Grade (Using a grade system) |
| Essay |
| Oral examination |
| Oral examination |
|
Recommended literature
|
-
Atkins, P.W. Physical Chemistry. Oxford.
-
I.D.Wilson, editor. Literatura: Encyclopedia of Separation Science, I. D. Wilson, editor, Academic Press, New York, 2000. Academic Press, New York, 2000.
-
Kodíček, M., Karpenko, V. Biofyzikální chemie. Praha, 1997.
-
Nováková, L., Douša, M. Moderní HPLC separace v teorii a praxi I.. Hradec Králové, Klatovy. ISBN 978-80-260-4243-3.
-
Nováková, L., Douša, M. Moderní HPLC separace v teorii a praxi II.. Hradec Králové, Klatovy. ISBN 978-80-260-4244-0.
-
Prosser, V. a kol. Experimentální metody biofyziky. . Praha, 1989.
-
Štulík, Karel. Analytické separační metody. 1. vyd. Praha : Karolinum, 2004. ISBN 80-246-0852-9.
|