BIOCHEMICAL AND BIOMOLECULAR TECHNIQUES WITH LABORATORY

The student will acquire skills related to the techniques for the analysis of the biochemical aspects of prokaryotes and eukaryotes and cell culture techniques, even on a large scale; for the study of the structure, functions and analysis of biological macromolecules and cellular processes.

Oral lectures integrated by laboratory activities. 

Should teaching be carried out in mixed mode or remotely, it may be necessary to introduce changes with respect to previous statements, in line with the programme planned and outlined in the syllabus.

 Learning assessment may also be carried out on line, should the conditions require it.

Knowledge acquired during the three-year degree courses in chemistry, biology and biotechnology

Classes are held during the second year, first semester, six to eight hours a week.

Cell cultures: primary cell cultures and cell lines.

Cell cycle analysis, cell proliferation, vitality and cell death assays.

Chain Polymerization Reaction (PCR), cDNA synthesis by reverse transcription. Bioinformatics parameters and softwares for PCR Primer Design (Primer 3 NCBI).

Qualitative and quantitative analysis of transcripts.

Evaluation of polymorphisms by quantitative PCR or "real-time PCR"

Digestion reactions by restriction enzymes. Reactions by modification enzymes.

Extraction, Purification and qualitative and quantitative determination of DNA, RNA  and proteins

Electrophoresis of nucleic acids and proteins.

Hybridization techniques. Southern blot, Northern blot and Western blot.

DNA or RNA sample preparations for global genome analysis and microarrays

Nucleic Acids Sequencing: Sanger and Next Generation Sequencing

Analysis of epigenetic modification (methylation)

Transient and stable transfection techniques and silencing of gene expression by "RNA interference"

Recombinant DNA technology and cloning: Preparation of a recombinant plasmid vector; Bacterial transformation and extraction of plasmid DNA

Enzymatic activity tests in biological samples.

Databases.

Biochemical and biomolecular laboratory techniques in medical molecular diagnostics.

Laboratory Experiences

Esp 1 human Cell lines

Esp 2 Nucleic Acids quantitative assay

Esp 3 DNA extraction from human cell cultures

Esp 4 Electrophoresis in agarose gel

Esp 5 cDNA synthesis

Esp 6 Polymerase Chain Reaction (PCR)

Esp 7 Digestion with Restriction Enzymes

Exp 8 Quantitative PCR / qRT Allelic Discrimination

Esp 9 Transfection and Silencing

1) Metodologie Biochimiche e Biomolecolari,  a cura di Maccarone M. e colleghi, Zanichelli editore

2) Biologia molecolare Autori: Francesco Amaldi, Piero Benedetti, Graziano Pesole, Paolo Plevani, II eds Casa Editrice Ambrosiana (2014) – Zanichelli

3) Metodologie biochimiche. Principi e tecniche per l'espressione, la purificazione e la caratterizzazione delle proteine Autori : M. Carmela Bonaccorsi di Patti, Roberto Contestabile. Editore: CEA

4) Additional  material supplied by  the teacher

Oral Examination. Verification of learning can also be carried out electronically, should the conditions require it.

The evaluation of the knowledge acquired takes place through an oral exam. The grade is expressed on a scale of thirty, up to a maximum of 30/30 cum laude.

The following parameters will be taken into account to assign the final grade:

• Score 29-30 with honors: The student demonstrates in-depth knowledge of the topics, promptly and correctly integrates and critically analyzes the situations presented, independently solving even very complex problems. He has excellent communication skills and competently masters the scientific language adopted.

• Score 26-28: The student has a good understanding of the topics, is able to integrate and analyze the situations presented critically and logically, explains the topics clearly using appropriate scientific language.

• Score 22-25: The student has a fair understanding of the topics, even if limited to the main areas. He is able to integrate and critically analyze the situations presented, although not always in a linear way, and presents the topics quite clearly with moderate linguistic competence.

• Score 18-21: The student has minimal knowledge of the topics, has a modest ability to integrate and critically analyze the situations presented and presents the topics clearly enough, although his/her linguistic competence may be poorly developed.

• Exam not passed: The student does not have the minimum required knowledge of the fundamental contents of the course. The ability to use a specific language is minimal or non-existent and the student is not able to independently apply the knowledge acquired.