Knowledge of methodologies and methodological schemes for the study of biomolecules

Protein purification Ionic properties of amino acids and proteins. Isoelectric point. Sample preparation. Methods of cell disruption and production of the initial crude extracts. Protein solubilization methods. Saline swabs. Fractionation and precipitation techniques. Filtration, dialysis, sample concentration. Spectroscopic techniques. Properties of electromagnetic radiation. Light-matter interaction. States and processes involved in the phenomena of absorption, emission and decay. Absorption spectroscopy in ultraviolet and visible. Qualitative and quantitative aspects of light absorption. Colorimetric and spectroscopic methods applied to the determination of protein concentration. Spectrophotometers. Fluorescence and emission spectroscopy. Intrinsic and extrinsic fluorophores. Green fluorescent protein Spectrofluorimeters. Fluorescence resonance energy transfer (FRET). Circular dichroism. Chromatographic techniques. Principles of chromatography. The chromatogram. Parameters that determine the chromatographic performance. Van Deemter equation. Column chromatography: ion exchange, molecular exclusion, affinity, hydrophobic interaction and their applications. Purification of fusion proteins with Electrophoretic techniques. General principles and electrophoretic mobility. Support materials. Nucleic acid electrophoresis. Protein electrophoresis. SDS PAGE. Electrophoresis in native conditions. Protein coloring on gel. Protein blotting (western blotting). Isoelectrofocusing. Basics of: Capillary electrophoresis. Two-dimensional electrophoresis gel.

The final (written) exam will focus on all the topics of the program. The student will have to demonstrate that they understand and be able to use the fundamental concepts of each topic.