Module

Voraussetzungen

Inhalte

Students will isolate expression vector DNA using different protocols and reagents and will later on compare the influence of these different procedures on transfection efficiency again applying different gene transfer methodologies. Here the students will perform the complete set of steps necessary for eukaryotic cell transfection. These experiments form the basis for the central part of the course the genome editing. Students will use the CRISPR/Cas system to induce genome mutations and create site directed specific sequence alteration. The analysis of the cell physiological effects of these editings will be performed using cell culture experiments. In addition little projects will be given to students including the following methods: coupling of nucleic acids and proteins to fluorescence coded microbeads: detection of the protein targets by antibody detection; nucleic acids are detected on the bead surface via hybridization or PCR; expression and purification of recombinant proteins; design of primers for qPCR, PCR and PCR based cloning. Because the master course is integrated into the scientific research of the group experiments will be adapted accordingly.

Lernziele

It is the aim of this laboratory course to provide basic molecular biology techniques in the first place. Students should experience isolation and manipulation of DNA and RNA in a way that they understand the underlying principles of these fundamental methods. In a second step, students have to learn different techniques used to transfect pro- and eukaryotic cells as these are important methods in biotechnology. Therefore, the course provides the complete scenario from the preparation of DNA up to the functional analysis of different expression plasmids. Third, students will become familiar with viruses, as these biological agents are important "work horses“ in different areas of molecular biology. Working with microbeads to detect nucleic acids and proteins in a multiplex format provides the opportunity for state of the art bioanalytical methods. This part is complemented by genome editing experiments. Students will get in touch with a state of the art complex methodology and get hands-on experience. The seminar has the task of broadening the theoretical basis of the methods used in the practical course. Presentations and enhanced discussions will provide the stage for a fruitful enhancement of molecular biology knowledge. In addition to specialized knowledge (professional competences), the students will train social competences because of being stimulated for contributions and critical discussions. Methodical competences are developed as well because students have to train their practical lab skills and generic competences as they have to solve scientific problems. All these projects require search and understanding of relevant scientific literature, design of experimental protocols, documentation and a critical discussion of results.

Prüfungsleistung

• Practical laboratory work-preparation of experiment, performance, discussion, time management (50%) • Presentation and discussion in the seminar; duration depends on the topic (25%) • Lab report (including title, abstract, introduction, methods and materials, results, discussion, conclusion, and references; 15-20 pages) (25%)

Bemerkungen

"Recommended Placement in the Study Plan" • Biotechnology (Master of Science, 3 Semester Version): Master 1st year course • Biotechnology (Master of Science, 4 Semester Version): Master 2nd year course The module takes place as a 6 week block course.