Learning outcomes of the course unit
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
Centrifugative techniques: differential centrifugation and centrifugation on density gradients.
Protein extraction and purification.
Chromatographic techniques: ion exchange; gel-filtration; affinity chromatography. HPLC.
Electrophoretic techniques: principles and applications. SDS-PAGE, isoelectrofocusing, 2D-PAGE.
Immunochemical techniques: immunoprecipitation, ELISA, Western blotting.
Spectrophotometric techniques (absorption and fluorescence) and their applications.
Use of radioisotopes in biochemical research: Geiger-Muller detectors, liquid scintillation, autoradiography, phosphorimaging.
Enzymatic assays: continuous, discontinuous and coupled assays. Unit of activity.
Basic notions on mass spectrometry: electronic impact ionization systems, ESI, MALDI. Detectors based on magnetic deflection, quadrupole and time-of-flight detectors
Ninfa & Ballou, Metodologie di base per la biochimica e la biotecnologia, Zanichelli ed.
Reed, Holmes, Weyers & Jones, Metodologie di base per le scienze biomolecolari, Zanichelli
The course will include formal classroom lessons only.
Assessment methods and criteria
Learning of the subject matters by the student 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.