CYTOGENETIC AND MOLECULAR TECHNIQUES

Knowledge and understanding

Students will acquire theoretical and practical knowledge of the principal cytogenetic and molecular genetic techniques, including:

• in vitro cell culture;
• classical cytogenetic techniques (preparation of metaphase chromosomes);
• molecular cytogenetic techniques (fluorescence in situ hybridization, FISH);
• DNA and RNA extraction and analysis;
• PCR, RT-qPCR, and gene expression analysis techniques.

Students will understand the biological and methodological principles underlying these techniques and their applications in the professional practice of biologists, particularly in biomedical and diagnostic settings.

Applying knowledge and understanding

By the end of the course, students will be able to:

• perform basic laboratory procedures, including cell culture, DNA/RNA extraction, and PCR;
• operate the principal laboratory instruments;
• analyze and interpret experimental results;
• apply cytogenetic and molecular genetic techniques in professional biological practice.

Making judgements

Students will develop the ability to critically evaluate experimental data and laboratory methodologies, with particular reference to the selection of appropriate experimental protocols and the interpretation of results.

Communication skills

Students will be able to describe laboratory procedures and experimental results using appropriate scientific terminology, both in written and oral form.

Learning skills

Students will acquire the skills necessary to independently explore advanced methodologies in cytogenetics and molecular genetics, including their diagnostic applications.

The course includes:

• Direct Teaching: face-to-face lectures;
• Interactive Teaching: practical laboratory activities.

Laboratory activities include:

• experimental demonstrations;
• hands-on laboratory exercises;
• implementation of experimental protocols;
• analysis and interpretation of experimental results.

Laboratory activities are essential for the development of practical skills and are conducted in accordance with applicable laboratory safety regulations, contributing to the acquisition of the fundamental professional competencies required for the practice of biology.

Should the course be delivered in blended-learning or online mode, appropriate adjustments to the teaching methods may be introduced while maintaining consistency with the intended learning outcomes and course contents described in this syllabus.

Information for students with disabilities and/or specific learning disorders (SLD). In accordance with current legislation and to ensure equal learning opportunities, students requiring accommodations are encouraged to arrange an individual meeting with the course instructor in order to discuss any appropriate compensatory and/or dispensatory measures based on the learning objectives and their specific needs. Students may also contact the departmental CInAP (Centro per l'Integrazione Attiva e Partecipata – Services for Students with Disabilities and Specific Learning Disorders) representative.

Students are expected to have basic knowledge of genetics and cell biology, with particular reference to:

• the structure and function of DNA and RNA;
• genome organization;
• cell division.

Successful completion of the Genetics course is a mandatory prerequisite for enrollment.

If the number of applications exceeds the maximum enrollment of 25 students, admission will be based on the grade obtained in the Genetics examination.

Access to laboratory activities requires prior completion of mandatory workplace and laboratory safety training.

Attendance is compulsory, in accordance with the Academic Regulations of the Degree Programme, as participation in the laboratory activities is essential for acquiring the practical skills envisaged by the course.

1. Safety in Genetics Laboratories. Chemical and biological safety in genetics laboratories. Personal protective equipment (PPE) and collective protective equipment (CPE). Safe handling of chemical reagents and biological materials. Risk identification in experimental procedures. Hazard statements and precautionary statements. Disposal of hazardous waste and the European Waste Catalogue (EWC/CER) classification system.

2. In Vitro Cell Culture. Primary cell cultures and established cell lines; culture media; cell maintenance; cryopreservation.

3. Classical Cytogenetics. Preparation of metaphase chromosomes; chromosome staining and banding techniques; microscopic chromosome analysis.

4. Molecular Cytogenetics. Fluorescence in situ hybridization (FISH); DNA probes; probe labeling; dual-color and multicolor FISH; image acquisition and analysis.

5. DNA Extraction and Preparation. Extraction of genomic and plasmid DNA; protocols for prokaryotic and eukaryotic cells; automated extraction systems.

6. DNA Polymorphisms and PCR. Restriction enzymes; restriction fragment length polymorphism (RFLP); polymerase chain reaction (PCR); gel electrophoresis; introduction to short tandem repeats (STRs) and multiplex PCR.

7. Gene Expression Analysis. RNA extraction; cDNA synthesis; qualitative and quantitative RT-PCR (RT-qPCR); experimental design and data analysis.

·       Lecture notes and other teaching materials provided by the instructor .

Textbook

·      Bridger JM, Volpi EV – FISH: protocols and applications. Humana Press

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

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

 

The final examination consists of three parts:

• a written examination (20 multiple-choice questions and 2 open-ended questions);
• a practical laboratory examination;
• an oral examination.

The final examination is designed to assess:

1. students' knowledge and understanding of the topics covered during the course;
2. their ability to analyze and synthesize scientific information;
3. their ability to apply the methodological skills acquired during lectures and laboratory sessions.

The final grade is expressed on a 30-point scale.

Assessment of laboratory activities

During the laboratory sessions, assessment will take into account:

• regular attendance and active participation in laboratory activities
• the ability to apply theoretical knowledge to experimental procedures
• the ability to understand and correctly follow laboratory protocols

Mid-term assessments

Two written mid-term examinations are scheduled, each consisting of:

• 20 multiple-choice questions;
• 2 open-ended questions.

The assessments cover the following topics:

• First mid-term examination: Cell culture, classical cytogenetics, and molecular cytogenetics;
• Second mid-term examination: DNA and RNA extraction and analysis, DNA polymorphisms, and gene expression analysis.

Assessment is expressed on a 30-point scale and is based on:

• knowledge and understanding;
• analytical and critical thinking skills;
• ability to apply laboratory methodologies.

Students who successfully pass both mid-term examinations may, during the first available examination session, be exempted from the written examination and proceed directly to the practical and oral examinations.

Oral examination

During the oral examination, assessment will focus on the relevance and accuracy of the answers, the quality of the scientific content, the ability to integrate concepts from different topics, the appropriate use of scientific terminology, and communication skills.

Information for students with disabilities and/or specific learning disorders (SLD): In accordance with current legislation and to ensure equal opportunities, students requiring accommodations may request an individual meeting with the instructor to discuss appropriate compensatory and/or dispensatory measures, consistent with the learning objectives of the course and their specific needs. Students may also contact the departmental representative of CInAP (Centre for Active and Participatory Inclusion – Services for Students with Disabilities and Specific Learning Disorders) for further assistance.

 

·      Restriction enzymes

·      Quantitative PCR (qPCR)

·      Restriction fragment length polymorphisms (RFLPs) and their applications in biomedical diagnostics

·      Established cell lines and primary cell cultures

·      DNA probe labeling for in situ hybridization