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Lecturer(s)
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Husová Michaela, Mgr.
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Tolar Nikolas, Mgr.
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Šauman Ivo, prof. RNDr. Ph.D.
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Gahurová Lenka, Mgr. Ph.D.
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Panicucci Zíková Alena, doc. RNDr. Ph.D.
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Mozgová Iva, Mgr. Ph.D.
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Nguyen Petr, RNDr. Ph.D.
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Hospodářská Monika, Mgr.
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Bajgar Adam, Mgr. Ph.D.
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Korytář Tomáš, Mgr. Ph.D.
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Žurovec Michal, prof. RNDr. CSc.
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Sehadová Hana, Mgr. Ph.D.
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Krejčová Gabriela, RNDr. Ph.D.
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Jindra Marek, prof. RNDr. CSc.
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Tůmová Šárka
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Course content
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Students select 2-3 of the listed practical sessions: 1. qPCR expression analysis (Adam Bajgar) 2. Advanced Drosophila genetics - generation and analysis of mitotic clones (Tomáš Doležal) 3. Systemic RNA interference in insects (David Doležel) 4. Microinjections and immuno-fluorescence staining of mouse embryos (Alexander Bruce) 5. Fluorescence in situ hybridization (Petr Nguyen) 6. Flow cytometry and cell sorting (Tomáš Korytář) 7. Detection of RNA modifications by afinity gel electrophoresis (Zdeněk Paris) 8. 2D native/SDS electrophoresis combined with immunodetection (Roman Sobotka) 9. Expression and purufication of His-tagged proteins (Roman Sobotka) 10. Classical histological techniques and light microscopy (Hana Sehadová) 11. Detection of high molecular weight respiratory complexes using native PAGE and subsequent Western blot analysis (Alena Zíková) 12. Measurement of mitochondrial respiration by High-Resolution Respirometry (HRR) using the O2k-FluoRespirometer (Eva Doleželová) 13. Western blot in insect tissues and cell lines (Marek Jindra) 14. Next generation sequencing data processing (Petr Nguyen) 15. RNA-seq library preparation and data analysis (Lenka Gahurová, Iva Mozgová) 16. Transfection of insect cell lines (Michal Žurovec)
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Monitoring, Demonstration, Laboratory, Experiment
- Preparation for classes
- 72 hours per semester
- Class attendance
- 48 hours per semester
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Learning outcomes
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The aim of the course is to provide practical hands-on experience with selected advanced molecular biology techniques
During the course, students will gain deeper knowledge and understanding about the majority of molecular biology techniques and their applications. Students should be able to independently select optimal methods for solving particular biological problems. They should also be able to advise other students on selection of the best method, and on its advantages and risks.
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Prerequisites
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Students should understand principles of molecular biology techniques. They should have basic knowledge from cell biology, molecular biology, developmental biology and genetics.
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Assessment methods and criteria
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Student performance assessment, Analysis of student's work activities (technical works)
To pass the practical part of the course, students must attend two selected practical sessions.
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Recommended literature
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Sambrook, J., Fritsch, E. J. and Maniatis, T. (1989) Molecular Cloning, A Laboratory Manual (2. vyd. ), Cold Spring Harbor Press..
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Ochrana František. Methodology of Science: An Introduction.
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