TECNICHE BIOCHIMICHE E BIOMOLECOLARI

Students will acquire both theoretical/practical knowledge of basic biomolecular methodologies useful for experimental analysis and application skills to be used in the field of biomolecular disciplines. During the scheduled laboratory hours, students will acquire practical skills in the use of the studied biomolecular techniques, with particular attention to the applications of genetic engineering and the study of recombinant proteins.

Frontal lessons and laboratory exercises

Nucleic acid and protein handling and analysis:

Isolation of DNA, RNA and proteins from biological samples; examples and applications. DIGE. Blotting (Western, Eastern).

Techniques for the analysis of gene expression:
Semi-quantitative, competitive PCR, qPCR. CDNA and oligonucleotide microarrays; SAGE, Differential-display

Analysis of protein-protein interactions:
In vitro display techniques: Protein Arrays, Pull-down, mRNA display and Ribosome display.
In vivo display techniques: Co-Immuncipitation, Tandem Affinity Purification (TAP) system, Phage Display

Analysis of DNA/RNA-protein interactions:
Chromatin immunoprecipitation assay (ChIP and ChIP-on-chip). EMSA. Triple hybrid test in yeast.

Mutagenesis techniques:
Site specific or random mutagenic methods: cassette mutagenic, primer extension mutagenic, PCR mutagenic. Mutagenesis applied to the study of promoters: linker-scanning mutagenesis. PCR applications in the diagnostic and forensic fields.

Sequencing:
Automatic DNA sequencing. Pyrosequencing. Next Generation Sequencing: platforms for second and third generation sequencing.

- Biotecnologie molecolari, principi e tecniche. Brown TA, 2017, Zanichelli editore

- DNA ricombinante. Watson, Caudy, Myers, Witkowski, 2008 ed., Zanichelli editore

- Genomi 3. Brown TA, ultima ed., EdiSes editore