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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 substance separation, intermolecular interactions and properties of substances resulting from them. 3. Processing of complex biological material, basic separation methods (filtration, extraction, precipitation, salting out). 4. Electromigratory 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, applications to equilibria of oligomeric proteins and polydisperse samples of polymers. Isopycnic centrifugation. 9. Chromatographic methods - basic principles of chromatography, analysis of results, chromatography performed in isocratic mode, gel permeation chromatography - principles and examples of use. Liquid and gas chromatography. 10. Gradient chromatographic methods - ion exchange chromatography, hydrophobic chromatography, reverse 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 determinations of proteins, amino acid analysis, protein sequencing, determination of DNA concentration, PCR and qPCR method. 14. Immunological methods, blotting, ELISA, flow cytometry.
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Learning activities and teaching methods
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- Home preparation for classes
- 20 hours per semester
- Participation in classes
- 56 hours per semester
- Preparation for examination
- 44 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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| Simple experiments |
| Simple experiments |
| Practice exercises |
| Practice exercises |
| assessment methods |
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| Knowledge |
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| Analysis of seminar paper |
| Analysis of seminar paper |
| Oral examination |
| Oral examination |
| Grade (Using a grade system) |
| Grade (Using a grade system) |
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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 978-80-260-4243-3.
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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.
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Štulík, Karel. Analytické separační metody. 1. vyd. Praha : Karolinum, 2004. ISBN 80-246-0852-9.
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WILSON, I.D. (Ed.). Encyclopedia of Separation Science.. New York: Academic Press, 2000.
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