Learning objectives
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.
Prerequisites
Basic courses in Physics, Chemistry, Microbiology and Biochemistry.
Course unit content
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.
NMR spectroscopy.
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.
Full programme
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Bibliography
Bonaccorsi di Patti, Contestabile, Di Salvo. Metodologie biochimiche, Casa
editrice Ambrosiana
Reed, Holmes, Weyers & Jones, Metodologie di base per le scienze
biomolecolari, Zanichelli
Teaching methods
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 specific topics or problems; 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.
Other information
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