Students following this course will receive a systematic preparation on the main techniques used in the biochemistry laboratory for the identification, isolation and characterization of biological macromolecules.
Basic courses in Physics, Chemistry, Microbiology and Biochemistry.
Course contents summary
Cell homogeneization and fractionation. Biochemical buffers.
Centrifugative techniques: differential centrifugation and centrifugation on density gradients.
Protein extraction and purification.
Chromatographic techniques: gel-filtration; ion exchange; hydrophobic exchange; affinity chromatography. HPLC.
Electrophoretic techniques: principles and applications. Protein electrophoresis: native, SDS-PAGE, isoelectrofocusing, 2D-PAGE. Electrophoresis of nucleic acids.
Optical spectroscopy and its applications: absorption, fluorescence and circular dichroism.
Immunochemical techniques: immunoprecipitation, ELISA, Western blotting.
Use of radioisotopes in biochemical research: Geiger-Muller detectors, liquid scintillation, autoradiography, phosphorimaging.
Enzymatic assays: continuous, discontinuous and coupled assays. Unit of activity.
Mass spectrometry: ionization systemsand detectors (magnetic deflection, quadrupole, time-of-flight, orbitrap, tandem detectors). Biochemical applications of mass spectrometry,
Bonaccorsi di Patti, Contestabile, Di Salvo. Metodologie biochimiche, Casa editrice Ambrosiana
Reed, Holmes, Weyers & Jones, Metodologie di base per le scienze biomolecolari, Zanichelli
The course will include formal classroom lessons only.
Assessment methods and criteria
The knowledge and understanding acquired by the student, as well as the ability to apply such a knowledge in practice, will be assessed through a final written test. The test will encompass both a section with multiple-choice quizzes and a section with open questions.
The multiple-choice quizzes will mainly serve to assess the acquired knowledge and understanding. As for open questions, the student will be required to write a text concerning a specific topic or problem; this will serve to assess his/her ability to apply knowledge and understanding, together with the capacity of making judgments and of communicating ideas and concepts with clarity and property of expression.