BIOCHEMICAL METHODS FOR THE ENVIRONMENT
Basics of Biochemistry.
Learning will be assessed through an oral exam, lasting about half an hour per student. Before the end of the course, articles and documents will be uploaded in e-learning, from which the student can choose the one to present during the exam. The student will have to prepare a presentation, in which he/she i) will describe the techniques used in the article, highlighting the theoretical bases learnt during the course, and ii) will critically discuss the relevance of the experimental results in the environmental field (also weaknesses or criticalities).
Evaluation criteria: the maximum mark will be 30.
Precision and significance in the theoretical contents and the ability to present, argue and summarize the contents with an appropriate language will be taken into account.
The correctness of the content and terminology is considered a fundamental requirement for passing the exam.
The ability to critically analyze the methodologies used and the results obtained will be taken into consideration to reach the maximum mark.
The aim of the course is to give students the theoretical basis of biochemical and molecular biology methodologies relevant to the figure of the modern environmental scientist.
At the end of the course, the student will be able to:
1) Understand the theoretical basis of the most common techniques of biochemistry and molecular biology
2) Understand scientific articles and normative documents that apply biochemical methodologies in the environmental field.
The course (6 CFU) is divided as follows:
- Introduction: biological samples to be analyzed and characterized. In vitro and in vivo models.
- Cell disruption and tissue homogenization; separation of organelles;
- Fractionation techniques: protein solubility and precipitation techniques. Centrifugation and ultracentrifugation. Filtration, dialysis, sample concentration. Freezing and freeze-drying.
- Chromatographic techniques: Column chromatography: ion exchange, hydrophobic interactions, gel filtration, affinity, partitioning, adsorption, HPLC.
- Electrophoretic techniques (of proteins and nucleic acids), SDS-PAGE, isoelectric focusing, two-dimensional electrophoresis. Specific colors.
- Immunochemical techniques: antibodies, Western blot, ELISA, Immunoprecipitation.
- Use of spectrophotometric techniques. Commonly used colorimetric assays. Methods of protein dosage and evaluation of enzymatic activity.
- Use of spectroscopic techniques in biochemistry (circular dichroism, fluorescence).
-Methods to produce recombinant proteins.
- Nucleic acid analysis techniques. Focus on the Polymerase chain reaction.
-Outlines of bioinformatics.
- The -omics techniques: genomics, transcriptomics, proteomics. Systematic proteomics and methodological basis of the proteomic approach: one- and two-dimensional protein electrophoresis; mass spectrometry coupled with database analysis. Proteomic study of post-translational modifications. Differential proteomics (SILAC, ICAT and iTRAQ techniques) and functional proteomics. Introduction to metabolomics.
-Hints of Systems biology
- journal club (1 CFU)
Classroom lectures (48 hours). The didactic material (presentations, links, scientific articles) will be made available to students before each lesson through the e-learning platform. With the aid of images, diagrams, graphics, the student will be guided to understand the techniques and approaches commonly used in biochemistry and molecular biology laboratories. Ample space will be devoted to the analysis of scientific articles or regulatory texts that apply biochemical methodologies in the environmental field, so that the student can appreciate the practical-applicative implications of the course and its usefulness for the training of a modern environmental scientist.
The teacher encourages the participation of students, through moments of discussion and shared analysis of the topics presented. In particular, to request clarification as soon as any topic is not clear, so as not to compromise the understanding of subsequent topics. Therefore, during the explanation, students are invited to ask questions and request for clarification or further information.
Attending students are required to proceed with the independent study of the reference texts, following the detailed program provided at the beginning of the course.
Non-attending students are required to study the texts indicated in the bibliography; they will be able to access the dedicated e-learning page to consult the materials produced by the lecturer, like the attending students. Furthermore, non-attending students who wish to have clarifications are invited to contact the lecturer for an appointment during reception hours.
The lecturer is available for further information and explanations, every day from Monday to Friday from 9 to 18, by appointment, by writing an email to tiziana.alberio@uninsubria.it
The meeting can take place in person (third floor office, via Manara, 7, Busto Arsizio) or using the Teams platform, individually or in small groups, if more than one student requests the same clarifications.