LABORATORY OF BIOCHEMICAL METHODOLOGIES

Course objectives

General skills. At the end of the course and after passing the exam, the student will have acquired the knowledge and skills in the areas below. In general, it will be able to: explain the principles and the applications of the most common biochemical methodologies and set up an experiment starting from the consultation of the literature up to its execution on the laboratory counter. On the basis of the acquired knowledge, the student will have the ability to interpret and explain the results of the biochemical experiments through the interpretation and discussion of the experimental results. Students' critical and judgmental skills will be developed through numerous classroom exercises in which numerical exercises and database research will be carried out, and several practical laboratory experiences (starting from cloning of PCR-produced DNA fragments to purification and characterization of proteins with enzymatic activity). Communication skills will be exercised during the theoretical and practical lessons, which will include moments of open discussion. In the future, the student will be able to count on the knowledge and skills herein acquired for work in analysis and research laboratories. Specific skills. a) Knowledge and understanding of the following: - methodologies concerning recombinant DNA (PCR, restriction, ligation); - preparative methods for the purification of proteins (centrifugation, chromatography); - analytical methodologies for the study of DNA and proteins (electrophoresis); - spectrophotometric techniques applied to the study of proteins - enzymatic kinetics; b) Ability to apply knowledge and understanding: - ability to apply the appropriate techniques to specific experimental problems, starting from scientific literature and database research; - ability to design and carry out biochemical experiments concerning structure and function of DNA, proteins and enzymes; - ability to interpret and discuss experimental results and to explain biological phenomena with biochemical argumentations; c) Autonomy of judgment: - to be able to solve autonomously both experimental and numerical problems; - to know how to select and evaluate the most appropriate techniques to solve a specific experimental problem; d) Communication skills: - to be able to illustrate and explain biochemical phenomena with appropriate terms and logical rigor; - to be able to describe the principles behind the main biochemical techniques; - to know how to discuss and describe the experimental results in a critical key; e) Learning ability: - acquisition of the fundamentals cognitive tools to be able to independently continue the study of biochemistry, both from a theoretical and a practical point of view; - acquisition of basic knowledge to progress autonomously in other related biological disciplines.

Channel 1
ANGELO TOTO Lecturers' profile

Program - Frequency - Exams

Course program
The course will be divided into two parts. The first part will be dedicated to the study of the main methodologies and techniques used in biochemistry and molecular biology laboratories. In the second part, topics related to bioactive peptides will be addressed, including techniques for their characterization, case studies, and biochemical/nanotechnological approaches for their application in the biomedical field. Lesson topics: The lesson topics may vary slightly from the outline below, depending on the level of preparation and interest of the group of students participating in the course. • General approach to biochemical research. • Methods for the expression of recombinant proteins. • Cell fractionation and purification of macromolecules. • Centrifugation techniques and solubility-based methods. • Chromatographic techniques. • Electrophoretic techniques. • Enzyme assays. • Spectrophotometric techniques. • Protein crystallization. • Use of the internet in biochemical research. • Bioactive peptides. • Techniques for the characterization of bioactive peptides. • Biochemical and nanotechnological approaches for the application of peptides in the biomedical field. • Use of fluorescent probes.
Prerequisites
1) ESSENTIAL: availability for the mandatory practical laboratory experiences and aptitude for group work; 2) IMPORTANT: students must have followed the courses of Biological Chemistry and Molecular Biology; the student should ideally have passed the related exams.
Books
• Bonaccorsi, Contestabile, di Salvo - Metodologie biochimiche – Zanichelli (2019), seconda edizione • Mauro Maccarrone - Metodologie biochimiche e biomolecolari. Strumenti e tecniche per il laboratorio del nuovo millennio. Zanichelli 2019 • Un libro di testo di Biochimica (Stryer, Voet&Voet, Leningher) • Lezioni su PowerPoint rese disponibili dal docente- Dispense e appunti di laboratorio
Teaching mode
The course will be carried out through theoretical lectures (24 hours), theoretical classroom exercises (10 hours) and practical laboratory experiences (26 hours). Lectures (24 hours): 1) Explanation of the topics covered by the program through slides and audiovisual material. This educational model is aimed at providing the theoretical knowledge of most common biochemical methodologies. 2) Open discussion of the topics of the lessons, during which the students are called to intervene, which have the purpose of developing the skills of communication, criticism and judgment. Theoretical exercises in class (10 hours): 1) Stoichiometry exercises on the preparation of buffer solutions and sample dilutions (2 hours). Particular attention will be paid to the understanding of the importance of buffer solutions and to the preparation of solutions in general 2) Exercises on nucleotide and protein sequences retrieving from databases (2 hours) 3) Exercises on cloning strategies of cDNA fragments into vectors (4 hours) 4) Compilation of a purification table (2 hours) Laboratory experiences (28 hours): 1) Preparation of buffer solution and broth by using the analytical balance and pH meter 2) Purification of a recombinant protein and analysis on SDS-PAGE (8 hours) 3) Preparation of model lipid membranes (8 hours) 4) Fluorescence assays (8 hours) The laboratory experiences, which are mandatory, are designed to put into practice the theoretical concepts studied in class, to develop the ability to plan an experiment and interpret it, to increase critical and judgmental skills
Frequency
Not mandatory
Exam mode
The exam includes a written test which, if sufficient, will allow the student to pass it. There will be the possibility to do an oral test to improve the result of the written test.
Bibliography
Reference bibliography will be provided together with lectures material
Lesson mode
The course will be carried out through theoretical lectures (84 hours) Lectures (48 hours): 1) Explanation of the topics covered by the program through slides and audiovisual material. This educational model is aimed at providing the theoretical knowledge of most common biochemical methodologies. 2) Open discussion of the topics of the lessons, during which the students are called to intervene, which have the purpose of developing the skills of communication, criticism and judgment.
BRUNO CASCIARO Lecturers' profile
  • Lesson code1019197
  • Academic year2025/2026
  • CourseBiology
  • CurriculumBiotecnologico cellulare
  • Year3rd year
  • Semester2nd semester
  • SSDBIO/10
  • CFU6