Over the last few years, research has faced an impressive advancement in experimental methods. This course intends to expose the students to a large number of techniques in order to build a solid knowledge on the wide variety of available experimental tools. The course aims to equip students with a detailed understanding and knowledge of the molecular principles underlying the advanced techniques currently used in research laboratories. Students will acquire a deep knowledge on the molecular strategies adopted to purify and sequence nucleic acids. The course will provide the theoretical principles on which the techniques discussed in class are based, and the practical and technical details on the main biochemical techniques such as electrophoresis, chromatography and in vitro techniques that use antibodies. Students will also develop a better understanding of the experimental strategies applied to genome manipulation.
Risultati apprendimento attesi
Students will benefit from this course by becoming competent in identifying the most appropriate techniques to use for their research purposes during their Master-thesis internship. Exposing the students to a large number of techniques, currently used in research, will help them to build a solid background toward the most basic methodological tools, acquiring the competence to identify the most appropriate experimental strategy to use for their research purposes. Students will learn both the theoretical principles on which the techniques discussed in class are based, and the practical and technical details on the main biochemical techniques such as electrophoresis, chromatography and in vitro techniques that use antibodies. Students will also develop a better understanding of the experimental strategies applied to genome manipulation.
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The program related to Genetics is aimed to illustrate the main methods used for the extraction and purification of nucleic acids, including genomic DNA, plasmid DNA and RNA. Moreover, the principles and procedures of PCR, RT-PCR and qRT-PCR assays are explained. The traditional methods of DNA sequencing, such as Sanger-Coulson and Maxam-Gilbert are discussed together with the Next-Generation sequencing approaches starting from the Solexa-Illumina, going through Ion current Ion 530 and Pacific Biosciences, to most the recent Oxford Nanopore Minion methods. The program related to Biochemistry will deal on strategies for protein expression in heterologous systems, spectroscopic techniques (absorbance and fluorescence), protein electrophoresis (SDS and Native PAGE and isoelectrofocusing), protein purification by chromatographic techniques, antibody structure and function and antibody-based techniques (western blot, Elisa). The program related to Molecular and Cellular Biology will deal with the methodologies used to introduce foreign nucleic acids in bacteria (transformation) as well as in cell cultures (transfection) and animal models (transgenic) and how to create a particular gene deletion (knock-out) or in animal models.
A revision of the topics covered by the basic molecular biology, biochemistry and biochemical methods classes included in the bachelor curriculum, before attending the course, is highly recommended.
Powerpoint-assisted frontal lectures with frequent, critical discussions on experimental design, protocols and specific case studies. Attendance is strongly suggested. Lecture slides and other reading/audiovisual materials will be made available on the ARIEL course website.
Materiale di riferimento
Students can refer to their Molecular Biology texts for the general aspects of nucleic acids and to books of Biochemistry (e.g. Stryer Lehninger, Ed. MacMillan) and of Principle and Techniques for Biochemistry and Molecular Biology (e.g. Wilson and Walker, Ed. Chiron Media (Wallingford, Regno Unito).
Modalità di verifica dell’apprendimento e criteri di valutazione
Evaluation of the student's performance will be based on an oral examination regarding the methodologies covered during the lessons. This part will contribute to the final grade with a maximum grade of 27/30 (90%). Furthermore, student performance during an exam session during which he/she will present and discuss a recent paper on advanced methodologies; this part will contribute to the final grade with a maximum grade of 3/30 (10%).