INGLESE

PCR and real-time PCR: theory and practice. Expression vectors: theory and practice. Transfection of
mammalian cells and application. Mouse models. RNA silencing: theory and application. Practice with
main bioinformatics tools for manipulation of nucleic acids

1-“Recombinant DNA. Genes and Genomes- A short course” JD Watson, AA Caudy, RM Myers, JA
Witkowski

2-Handouts for laboratory experiences

3- Slides of lessons on Department web site

Main aims of the course:
-Knowledge of advanced techniques in Molecular Biology.
-Practice with some molecular biology techniques commonly used in a wet laboratory
- Development of attitude to use the knowledge of molecular mechanisms to envisage innovative techniques.

Knowledge and skills provided by a basic course of Molecular Biology

Theoretical lessons and laboratory experiences in 8 lessons (one lesson: 4 hours) for a total of 32 hours.
Each lesson consists of:
2 hours, frontal lesson (basic theory)
2 hours, laboratory experiences in the laboratory of Experimental Biology

Attendance to laboratory experiences are compulsory.

Oral examination.
The oral examination will ascertain the comprehension of the different topics, and the ability to immagine experimental approaches to study, at molecular level, a simple biotechnological issue.

The student will be supported in his/her own studies by slides and handouts published on the web site of Department.

The teacher will be available for students as indicated, and upon request via email.

Theoretical lessons: PCR. Principles and applications. RT-PCR, 5’-RACE, 3’-RACE, real-time. Expression vectors. Expression of recombinant proteins in prokaryotes and eukaryotes: basic principles and applications. Transfection of mammalian cells: Principles and application. Mouse models for human diseases: basic principles and applications. RNA silencing: Molecular mechanisms and Application in molecular biotechnology. RNA regulatory network: basic principles and applications.

Laboratory experience: Project: "Production of recombinant K-RAS oncoprotein" throughout the following steps: Practice with the main bioinformatics tools for manipulation of nucleic acids. Searching for sequences in databank, multiple sequence alignment, design of PCR primers, restriction map and virtual cloning. Wet lab: RNA purification from human cell lines; Isolation of K-ras transcript by RT-PCR; cloning of K-ras cDNA in the expression vector pcDNA3.1; cell culturing.
Additional experience: Design of a detection kit for a virus. Design of an expression vector for the production of Spike protein. Design of a reporter vector for miRNA target validation.

English