BIC-10306 Practical Biological Chemistry

Vak

Studiepunten 6.00

Language of instruction:

Dutch

Contents:

This introductory practical course provides basic knowledge and skills in molecular biology and biochemistry. Lectures provide the theoretical background of the applied techniques and the experiments to be performed. Subjects are: techniques of gene cloning, structure and function of nucleic acids and proteins, structure and expression of eukaryotic genes, electrophoresis, PCR, hybridization, spectrophotometry and fluorescence. The practical consists of two parts: one molecular-biological and the other biochemical. The molecular-biological part is dedicated to: isolation and analysis of DNA, RNA and proteins, both from prokaryotes (E. coli) and eukaryotes (tomato); construction of a physical map of a plasmid by enzyme digestions and electrophoresis; cloning; detection techniques like PCR and the use of computer programs and internet in the analysis of structure and function of DNA (bio-informatics). The gene encoding one of the Rubisco proteins will serve as a model in part of the experiments. The biochemical part comprises the following subjects: quantitative clinical-chemical enzyme assays (uric acid quantitation, enzymatic assay for a heart-attack), detection of iso-enzymes with iso-electric focussing (tissue analysis), following metabolic processes in vivo using fluorescence (regulation of glycolysis in yeast cells), a study of the reaction mechanism of enzymes with enzyme kinetics and computer graphics, design of an immunochemical hormone assay and design and execution of a fluorimetric enzyme activity assay.

Learning outcomes:

After successful completion of this course students are expected to be able to:
- describe general genome structure and gene organization, transcription and RNA processing in eukaryotes;
- explain and apply basic principles of DNA cloning, hybridization and PCR;
- apply basic techniques in molecular biology, such as DNA and RNA isolation, PCR, restriction enzyme digestions and analysis, agarose gel electrophoresis, cloning;
- use DNA- and protein analysis software and perform simple database searches on the internet;
- understand the principles of spectrophotometry, fluorometry, immunological methods and electrophoresis and the use of these techniques in biochemical analyses;
- design and perform enzymatic assays;
- calculate and interpret independently the results of their experiments.

Activities:

Attending lectures and participation in the practical course. During the practical course, experimental results are elaborated and a report is written. Students receive tasks that should be solved during the course by Internet searches.

Examination:

The final mark is based on the results of an exam, laboratory skills and on the quality of the report of the practical part.

Literature:

A manual for the practical course Biological Chemistry is available at the secretary of the Laboratories for Biochemistry and Molecular Biology. The theoretical background of this course can be found in:
- Berg, Tymoczko & Stryer: 'Biochemistry' (7th ed, WUR shop);
- Griffiths et al.; 'Introduction to genetic analysis' (9th ed).