CYTOGENETIC AND MOLECULAR TECHNIQUES

The CYTOGENETIC AND MOLECULAR TECHNIQUES consents to the students the acquisition of theoretical and practical knowledge and understanding of the main laboratory techniques for cytogenetic and molecular investigations, such as cell cultures, the fluorescent in situ hybridization, analysis of DNA / RNA, PCR, RT-PCR, electrophoresis including the use of texts and teaching materials in English. The student also acquires the ability to perform the above methods and use the main instruments in use in various fields of work on the profession of Biologist. The student, at the end of the course, will acquire the skills to perform basic experimental procedures and analyze the results.

Basic methods for DNA preparation. The preparation of genomic and plasmidic DNA. The standard experimental protocols. The automated systems for DNA preparation.

The DNA polymorphisms. Restriction enzymes. PCR. The RFLP and their use in biomedical diagnostics. Genotypic through a survey with RFLP. Analysis by means of elechtrophoresis. Multiplex PCR and STR.

Gene expression analysis. RNA extraction from cells and tissues. Preparation of cDNA and methods of use. Methods of qualitative and quantitative analysis of a RNA. The quantitative PCR. Design of a quantitative RT-PCR and analysis of results.

The in vitro cell cultures. The cells in primary culture and cell lines. The culture media. Maintenance mode of cell cultures. The cryo-preservation and freezing procedures / cell thawing.

Molecular cytogenetics. The fluorescence in situ hybridization (FISH). The DNA probes: preparation methods, labeling and use. Labeling by antigenic molecules and fluorescent molecules. The dual-color and multicolor FISH. Image acquisition by fluorescence microscopy and analysis of results.

Educational material will be provided by the teacher during class.

Moreover, some books could be used for further details, such as:

Bridger JM and Volpi EV. Fluorescence in situ hybridization (FISH): protocols and applications. Humana Press.

Campbell LJ. Cancer cytogenetics: methods and protocols. Humana Press.

Brown TA. Biotecnologie molecolari: principi e tecniche. Zanichelli.