Students are provided with knowledge on the main techniques used in biochemical laboratories for the purification and characterization of biological molecules. In particular, the course focuses on learning the basic theoretical and practical principles of spectroscopic, chromatographic and electrophoretic techniques.
Course Prerequisites
Knowledge of basic chemistry and biochemistry
Teaching Methods
Lecture, where Power Point projections are used.
Assessment Methods
Oral examination. The exam consists of an individual oral test, where the skills acquired in relation to the course contents will be verified.
Texts
Didactic material provided by the teacher
Contents
Basic principles: Mass, volume, solutions, concentration, pH. Preparation of solutions, buffers and use of the basic instrumentation of a laboratory. Deionized water and MilliQ. Spectroscopic techniques. Main theoretical principles: properties of electromagnetic radiation, interaction with matter. UV and visible spectroscopy; theoretical principles, Lambert-Beer law, instrumentation, applications. Spectrofluorimetry, Colorimetry. Chromatographic techniques: General principles. Low pressure and high pressure chromatography (HPLC) Molecular exclusion chromatography (Gel filtration) Ion exchange chromatography Affinity chromatography Criteria for choosing a chromatographic system. Electrophoretic techniques: General Principles Support materials: Agarose, Polyacrylamide. Protein electrophoresis: SDS-PAGE, electrophoresis in native conditions, Isoelectrofocusing (IEF), Two-dimensional electrophoresis (2-D PAGE) Staining and revelation methods, Western blotting. DNA electrophoresis on agarose gel. Capillary electrophoresis (outline).
