8S130 - Molecular and cell biological techniques

Contents

The main goal of this course is to teach several important molecular and cell biological techniques. Students will work on several practical assignments that together form one research project. This project starts at the laboratory of protein engineering. An expression vector encoding for a GFP fusion protein that is important for cell structure will be transformed in E. coli and amplified. After purification the plasmid DNA will be characterized using restriction analysis. In the laboratory for Cell&Tissue engineering, the plasmid will then be transfected into a mammalian cell line. The expression and cellular localization will be studied using fluorescence microscopy. Expression of the fusion protein will also be analyzed using Western blotting. Each student will work individually, an assistant will be available in both labs to assist students with their practical work. In addition to the practical work some lectures will be given on safe microbiological practices, regulations regarding GMOs, cell culturing and fluorescence microscopy. At the end of the project students will thoroughly discuss their work on basis of their lab journal which has been kept up-to-date during their practical work in a correct manner. Knowledge on safe microbiological techniques will be tested by solving a few case-studies. After a succesful defense students will get a so-called VMT-certificate that allows them to work with GMOs in future research projects.

Learning objectives

Practical training in some important molecular biology and cell biology techniques, to obtain both practical skills and theoretical knowledge in working safely with genetically modified organisms (SMT-certificate), learn to plan and carry out experimental work independently.

Lecturers

Responsible lecturer: M. (Moniek) de Liefde - van Beest.

Co-lecturers: M. (Maarten) Merkx, C.C. (René) van Donkelaar.