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Lecturer(s)
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Kopečný David, Mgr.
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Zalabák David, Mgr. Ph.D.
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Course content
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1. Inoculation and cultivation of E. coli cells bearing selected plasmids. Isolation of plasmid DNA from bacterial cells. Restriction cleavage of plasmid DNA. Agarose electrophoresis. DNA quantitation. 2. Detection of recombinant DNA in food by PCR. Isolation of DNA from flour. Setup and evaluation of the PCR reaction. 3. Transformation of yeast S. cerevisiea using a shuttle vector and integration vector bearing the gene Atckx2. Selection of transformants and comparison of transformation yield. Evaluation by measuring CKX activity. 4. Work with bacteriophage cDNA library. Cultivation of the phage, DNA isolation. Determination of the titre of the library. Detection of a low abundant and a house-keeping gene. 5. Isolation of RNA from plant material, reverse transcription and RT PCR.
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Learning activities and teaching methods
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Dialogic Lecture (Discussion, Dialog, Brainstorming), Demonstration, Laboratory Work
- Preparation for the Course Credit
- 30 hours per semester
- Homework for Teaching
- 70 hours per semester
- Attendace
- 100 hours per semester
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Learning outcomes
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Practising of standard molecular biology methods.
Apply methods and procedures used in molecular and cell biology
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Prerequisites
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Subject KBC/MBIOG
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Assessment methods and criteria
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Student performance, Systematic Observation of Student
Student is required to complete all experimental tasks, obtain 80% of the points (based on the quality and integrity of the results) and submit a research report on each task. Before entering the practicals, it is compulsory to pass the examination KBC/MBIOG.
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Recommended literature
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Alberts B, Johnson A, Lewis J, Morgan D, Raff M, Roberts K, Walter P. (2017). Molecular Biology of the Cell, 6th edition. Garland Science.
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Green M.R., Sambrook J. (2012). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press.
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Šmehilová M, Dzurová L, Frébort I, Galuszka P. (2014). Laboratorní cvičení z molekulární biologie. elektroniká verze.
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