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Lecturer(s)
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Ingr Marek, doc. RNDr. Ph.D.
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Course content
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1. Introduction to separation methods, historical overview. 2. Principles of separation of substances, intermolecular interactions and properties of substances resulting from them. 3. Processing of complex biological material, basic separation methods (filtration, extraction, precipitation, desalting). 4. Electromigration separation methods - theoretical basis. 5. Capillary zone electrophoresis, isotachophoresis, isoelectric focusing - principles and applications. 6. Gel electrophoresis and special electromigration methods (DNA sequencing, MEKC). 7. Sedimentation methods as a tool for the analysis of natural substances. Sedimentation rate method. 8. Sedimentation equilibrium, application to oligomeric protein equilibria and polydisperse polymer samples. Isopycnic centrifugation. 9. Chromatographic methods - basic principles of chromatography, analysis of results, chromatography performed in isocratic mode, gel permeation chromatography - principles and examples of application. Liquid and gas chromatography. 10. Gradient chromatographic methods - ion-exchange chromatography, hydrophobic chromatography, reversed-phase chromatography, affinity chromatography - principles and examples of use. Thin-layer chromatography. 11. Detection methods in chromatographic and electromigration methods - basics of spectrophotometry and fluorimetry, detection in gas chromatography. 12. Refractometry, conductometry, basics of mass spectrometry. 13. determination of concentrations of biologically active substances - colorimetric determination of proteins, amino acid analysis 14. Immunological methods, blotting, ELISA, flow cytometry.
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Learning activities and teaching methods
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Lecturing
- Participation in classes
- 28 hours per semester
- Preparation for course credit
- 24 hours per semester
- Preparation for examination
- 24 hours per semester
- Home preparation for classes
- 14 hours per semester
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| learning outcomes |
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| Knowledge |
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| 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 |
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| 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 |
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| Knowledge |
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| Lecturing |
| Lecturing |
| Dialogic (Discussion, conversation, brainstorming) |
| Dialogic (Discussion, conversation, brainstorming) |
| Skills |
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| Methods for working with texts (Textbook, book) |
| Methods for working with texts (Textbook, book) |
| Individual work of students |
| Individual work of students |
| assessment methods |
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| Knowledge |
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| Grade (Using a grade system) |
| Grade (Using a grade system) |
| Analysis of seminar paper |
| Analysis of seminar paper |
| Oral examination |
| Oral examination |
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Recommended literature
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Prezentace z přednášky.
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ATKINS, P.W. Atkins´ Physical Chemistry. 10th Ed. New York: Oxford University Press, 2014. ISBN 9780199697403.
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Kodíček, M., Karpenko, V. Biofyzikální chemie. Praha, 1997.
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Nováková, L., Douša, M. Moderní HPLC separace v teorii a praxi I. Hradec Králové, Klatovy. ISBN 978-80-260-4244.
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Nováková, L., Douša, M. Moderní HPLC separace v teorii a praxi II.. Hradec Králové, Klatovy. ISBN 978-80-260-4244-0.
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Prosser V. a kol. Experimentální metody biofyziky. Praha, 1989. ISBN 80-244-0602-0.
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Štulík, Karel. Analytické separační metody. 1. vyd. Praha : Karolinum, 2004. ISBN 80-246-0852-9.
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