Learning outcomes of the course unit
Molecular Biology: the use of examples and practical case-histories to explain the possible applications of these techniques in the Biotechnology sector; fundamental concepts and notions of Molecular Biology.
Course contents summary
The stability, denaturation-renaturation, and melting-annealing of nucleic acids: Southern blot, Northern blot and "indirect labelling".
The structure/composition of nucleic acids: direct marking strategies; oligonucleotide synthesis and various forms of controlled DNA and RNA breakdown.
Restriction enzymes: structure, function, principal applications.
DNA replication and its biotechnological applications: the preparation of DNA probes; various types of PCR; molecular diagnosis based on PCR (human pathologies, viruses); enzymatic DNA sequencing (the Sanger method); "deep sequencing" by means of pyrosequencing.
DNA transcription: the use of “primer extension” to map transcripts; experimental set-up and interpretation of the results generated by analysis and protein-DNA interaction by means of “footprinting” and “electrophoretic mobility shift assay” (EMSA).
Retro-transcription: RT-PCR and its applications, the construction of cDNA gene libraries.
Site-specific recombination and its applications: Cre and FLP recombinase and the creation of conditional knock-out mutants, the targeted elimination of selection markers, and the site-specific activation of transgenes; molecular cloning using the Gateway system.
Watson J.D., Backer T.A., Bell S. P., Gann A., Levine M., Losick R., BIOLOGIA MOLECOLARE DEL GENE (V Edition), Zanichelli Editore, 2005
H. Lodish et al., MOLECULAR CELL BIOLOGY, Freeman, 2008
Specific course material provided for students