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Lecturer(s)
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Morávek Ondřej, Mgr.
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Beránek Martin, prof. PharmDr. Ph.D.
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Jankovičová Barbora, Mgr. Ph.D.
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Course content
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Selection of the gene of interest in the yeast genomic database, primer design. DNA isolation from the yeast Saccharomyces cerevisiae. Preparation of competent Escherichia coli cells. Amplification of a selected gene from yeast DNA by PCR. Control of PCR products by electrophoresis and their purification. Insertion of the PCR product into the vector using a kit. Preparation of selection media, inoculation of colonies into liquid medium. Mini-preparation of plasmid DNA. Restriction digestion of the resulting vector. Electrophoretic analysis of starting and final vector and restriction grafts. Preparation of RNA isolation aids. Isolation of RNA from blood. Electrophoretic and spectrophotometric characterization of the obtained RNA. Overall evaluation of results, including economic complexity of experiments.
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Laboratory work
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Learning outcomes
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The main goal of the course is to master the practical isolation of nucleic acids and their analysis, mastering the PCR method and interpretation of partial results.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Written examination, Home assignment evaluation
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Recommended literature
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aktuální návody na jednotlivá cvičení (portál STAG Upa).
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T. Ruml. Laboratoře z genového inženýrství. Praha: VŠCHT, 1997.
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