Aim of the course is to provide students with the basic knowledge of techniques typically applied in a biochemical laboratory. The formative aims are shared by the two parts of the course, that will describe dinstint, but strongly related topics. On completion of the course, the student should have acquired basic knowledge of the experimental procedures to be used in a biochemical and/or molecular biology laboratory.
Course Prerequisites
Basic knowledge of biochemistry and molecular biology. Knowledge of the general concepts about protein structure and function.
Teaching Methods
Frontal lectures, seminars on specific topics, guided lab tours for small groups and description of equipments and facilities for biochemical investigations.
Lecture slides and further information will be available on the KIRO platform throughout the course.
Additional support and teaching aids will be guaranteed exclusively to students belonging to specific categories defined by the University governance (working students, disabled, caregivers, etc.), who will not be able to attend the lessons in presence.
Assessment Methods
Oral examination. Discussion of a scientific article. During the examination, all topics indicated in the syllabus will be considered.The scores will be determined by the commission and expressed on a 30 points scale. Appropriate examination formats are provided for students who fall into the categories provided for Specific Learning Disorders (SLD) and Special Educational Needs (SED).
Texts
Principles and Techniques of Practical Biochemistry by K. Wilson and J. Walker. 5th Edition. Cambridge University Press. Course notes and material provided by the teacher and videos shared on the KIRO platform. The didactic materials provided by the instructor, in the form of PowerPoint slides and/or PDF files, are intended solely for the use of students enrolled in the 2024/2025 academic year of the Master’s course in Molecular Biology and Genetics, Molecular Life Science. These materials are designed to support the understanding of topics covered in each lecture. They are not a substitute for the recommended textbooks or original sources referenced by the instructor. Reproduction, distribution, or publication of any content (including text, audio, or video) in any form is strictly prohibited.
Contents
Part 1) Introduction to proteins. Cloning and expression of recombinant proteins. Protein extraction from animal/plant tissues and cell cultures. Primary purification processes. Chromatographic methods (IEC, SEC, HIC, affinity). Electrophoresis (native and SDS-PAGE, IEF). Spectrophotometry in protein assays and enzymatic assays. Immunoassays: antibody production. ELISA and Western Blotting. Fluorescence microscopy and flow cytometry. Use of radioisotopes in biochemistry. Part 2) HPLC, FPLC, gas chromatography, supercritical fluid chromatography. Two-dimensional electrophoresis, capillary electrophoresis. Application of spectroscopic techniques (UV, visible, fluorescence, mass, NMR, ESR) in proteomics studies. Mass spectrometry. Determination of the amino acid composition of a protein and study of its primary structure.