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Universitätsklinik für Neurologie Universitätsklinik für Neurologie

Mag.Dr. Theologos Michaelidis Mag.Dr. Theologos Michaelidis

519168

Embryonic Stem Cells: Culture, Genetic Modification, In Vitro Differentiation

UE 4

Block

semestral

keine Angabe

The course trains students in essential Gene Targeting and ES cell culture protocols and provides them with the scientific background required to make genetic alterations in mice. In parallel, they get practical experience with current in vitro differentiation protocols, particularly into the neuronal stages. The students are encouraged to design and perform the experiments by themselves, with our continuous support, such that at the end of the course they will be able not only to understand but also to apply this technology on their own work.

The course covers modern methods to retrieve and process genomic information using sequence databases as well as design of experimental strategies to generate conditional gene alterations. The students get substantial experience in handling macromolecular DNA (propagation of lambda bacteriophages, isolation of genomic DNA and restriction analysis), ES cell culture methods (preparation of mouse embryonic fibroblasts and feeders, freezing, thawing and culturing of ES cells, electroporation and antibiotic selection, karyotyping) and in vitro differentiation protocols. They also visit the animal house and make genotypic analysis of available knockout animals. Along with the experiments, relevant topics from scientific journals and books are discussed on a regular basis. The optimal number of students is 4. Methode Vorträge und praktische Übungen. The course requires a ~3 hrs daily attendance for 3 complete weeks. Each day starts with a discussion, which lasts approximately 30 to 40 min and includes the theoretical background of the experimental procedures, which will follow. 20 to 30 min before we finish, we discuss the protocols of the following day in order to estimate the time and steps that are required and make a convenient plan for all. Throughout the course the students keep detailed notes and record most experimental steps by photographing.

Vorträge und praktische Übungen. The course requires a ~3 hrs daily attendance for 3 complete weeks. Each day starts with a discussion, which lasts approximately 30 to 40 min and includes the theoretical background of the experimental procedures, which will follow. 20 to 30 min before we finish, we discuss the protocols of the following day in order to estimate the time and steps that are required and make a convenient plan for all. Throughout the course the students keep detailed notes and record most experimental steps by photographing.

- Gene Targeting: A Practical Approach (Practical Approach Series, 212) Oxford Univ. Press; Joyner AL ed. 2000. - Manipulating the Mouse Embryo: A Laboratory Manual. 3d ed. 2003. Nagy, Gertsenstein, Vintersten, Behringer; Cold Spring Harbor Laboratory Press.

The course will be held in English.
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