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.

PCR. Principles and applications. RT-PCR, 5’- RACE, 3’-RACE, real-time. Laboratory experience: Isolation of
human Dicer transcript by RT-PCR; Design of a detection kit for a virus.
Expression vectors. Expression of recombinant proteins in prokaryotes and eukaryotes: basic principles
and applications. Laboratory experience: cloning of Dicer cDNA in the expression vector pcDNA3.1; Design of an expression vector for the production of a viral recombinant protein.
Transfection of mammalian cells: Principles and application. Laboratory experience: cell culturing.
Mouse models. Production of transgenic mice for studying gene expression control: gene trap, enhancer
trap, cellular ablation. Production of knock-out and knock-in mice for studying molecular basis of human
diseases. Crispr-Cas9 system.
RNA silencing. Molecular mechanism. siRNA and miRNA. Application in molecular biotechnology.
Laboratory experience: design of a reporter vector for miRNA target validation.
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

English