Syllabus
During the first part of the course, students will first learn basic laboratory tasks such as preparation and sterilization of solutions, buffers and microbiological media. It also introduces the basics of sterile work, inoculation and cultivation of microorganisms on solid and liquid media and monitoring of the growth curve.
In the second part of the course, students will learn the methods needed to work with DNA, such as plasmid DNA electroporation into bacteria, its growth and preparation of bacteria, and electrophoretic verification of isolated plasmid DNA including restriction analysis. Plasmid DNA thus obtained will be used as a template for in-vitro cloning of the transformation cartridge using PCR with primers according to its own design. The transformation cartridges will then be transformed into a yeast strain and the success of the prepared gene fusion will be verified again using PCR with primers according to their own design.
The third part of the course is devoted to analyzing prepared strains. Genetically modified yeast strains will be analyzed using a fluorescent microscope, spectrofluorimeter and flow cytometry. For these analyses will be used instrumentation in BIOCEVu in Vestci near Prague.
Annotation
Two-semester Oxford-type practical course is focused on work with model organism - the yeast Saccharomyces cerevisiae. Each student will solve his own mini-project consisting of construction of a yeast strain carrying gene of interest fused with green fluorescent protein or other fluorescent protein in the genome and subsequent analysis of this strain.
The course is designed as an introduction to laboratory work with a real project. At each stage of the experimental work, students will learn the methods used for each step and subsequently will use the new know- how to get familiar with individual steps of preparing their own genetically modified yeast strain.
The course capacity is limited by the complexity of the methods used, but the low number of course participants allows to maximize individual approach and provides sufficient opportunities for discussion. Students are enrolled in the course on the basis of interest assessment test.