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Lecturer(s)
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Vrzalová Aneta, Mgr. Ph.D.
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Course content
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The techniques and principles of work with human carcinoma cell lines under sterile conditions. Cultivation of human liver hepatocellular cells HepG2, trypsinization and passaging of cells, determination of a number and a viability, a cell seeding. Transient transfection of cells with a reporter plasmid using a lipofection technique. Design of the pharmacological experiment, preparation of solutions of agonists and antagonists, treatment of transiently transfected HepG2 cells. Cell lysis permeabilization using surfactant, determining the catalytic activity of firefly luciferase using an automatic chemiluminometry. Calculation of IC50 values for the antagonist and agonist EC50.
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Learning activities and teaching methods
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unspecified
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Learning outcomes
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Introduction to the practical implementation of modern molecular biological techniques - work with cancer cell lines, reporter gene assay.
After completing the course the student is able to: - apply the assay in routine practice, in academic or research sector - Work under sterile conditions with human cancer cell lines - Work with technique of transient transfection by lipofection - evaluate of luciferase activity and the reporter gene assay
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Prerequisites
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unspecified
KBB/MTSB
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Assessment methods and criteria
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Student performance, Didactic Test
100% atendance at practicals, 70% test
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Recommended literature
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