CYTOGENETIC AND MOLECULAR TECHNIQUES
Academic Year 2023/2024 - Teacher: CONCETTA FEDERICOExpected Learning Outcomes
Course Structure
Classroom lessons and laboratory activities.
Should teaching be carried out in mixed mode or remotely, it may be necessary to introduce changes with respect to previous statements, in line with the programme planned and outlined in the syllabus.
Detailed Course Content
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.
Classical cytogenetics. Preparation of the metaphase chromosomes from cell cultures and human lymphocytes. Chromosome staining and banding techniques and observation with the microscopy.
Molecular cytogenetic. The fluorescence in situ hybridization (FISH). The DNA probes: preparation methods, labeling and use. The dual-color and multicolor FISH. Image acquisition by fluorescence microscopy and analysis of results.
Basic methods for DNA preparation. The preparation of genomic and plasmidic DNA. The different protocols of DNA extraction from bacteria and eukaryotic cells. 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.
Textbook Information
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.
Course Planning
Subjects | Text References | |
---|---|---|
1 | Le colture cellulari in vitro. | PDF 1: colture cellulari |
2 | Citogenetica classica. | PDF 2: Citogenetica |
3 | Citogenetica molecolare. | PDF 3: citogenetica molecolare |
4 | Espressione genica. | PDF 5: qRT-PCR. |