BIOCHIMICA APPLICATA E TECNOLOGIE BIOCHIMICHE PER LE BIOTECNOLOGIE

9 CFU - 1° Semester

Teaching Staff

DANIELE FILIPPO CONDORELLI - Module Applied Biochemistry - BIO/10 - 6 CFU
VINCENZA BARRESI - Module Molecular Diagnostic Technologies - BIO/12 - 3 CFU

Learning Objectives

Applied Biochemistry
Knowledge on structure, function and regulation of biological macromolecules and metabolic pathways.
Molecular Diagnostic Technologies
The Molecular Diagnostic Technologies have broad application in the biomedical field and the course aims to give the student the knowledge of methods and tools to be used for the detection of gene mutations and genomic alterations necessary for diagnostic, prognostic and predictive of response before the start of targeted therapeutic protocols

Course Structure

Applied Biochemistry
Classroom lessons and exercises.

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.
Molecular Diagnostic Technologies
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.

Detailed Course Content

Applied Biochemistry
1. organ and tissue biochemistry;
2. signal transduction pathways described at the level of enzymatic activities and protein interactions;
3. DNA damage and repair;
4. biochemical techniques for detection and characterization of proteins, nuclei acids and metabolites.
Molecular Diagnostic Technologies
Biological samples: types, blood and biological samples, processing and storage.

Extraction and purification of nucleic acids from human cells by chromatographic columns for gel filtration, ion exchange and affinity. Purification of nuclec acids with magnetic beads.

Quantitative assay for nucleic acids using spectrophotometric and fluorimetric methods.

Technologies based on the complementarity of the purine and pyrimidine nucleotides.

Synthesis of cDNA by reverse transcription.

Polymerase chain reaction (PCR). Qualitative PCR or End Point and quantitative PCR or Q-Real Time PCR. Programs for the design of primers to be used for PCR reactions.

Evaluation of gene expression in real time by the method of 2-ΔΔCT. for Allelic discriminationusing the Real Time PCR. Restriction and Modification enzymes.

Agarose gel electrophoresis for the separation of nucleic acids. Capillary electrophoresis for acid nucleic sequencing and analysis of microsatellite.

Protein extraction and analysis. Electrophoresis of proteins. Western blot.

DNA- and RNA-microarrays: Preparation of samples of DNA or RNA for the global analysis of the genome and transcriptome. Molecular analysis of the human karyotype microarray genomic photolithography-based probes to "Single Nucleotide Polymorphism-SNP" and "Copy Number Variation-CNV". Using algorithms and softwares for the interpretation of the data (CN state, Allele difference). Transcriptome analysis. Microarray expression to oligonucleotides obtained by photolithography synthesis. Array-CGH. Using algorithms and softwares for data interpretation (RMA and SAM).

Application of the "microarray" technology for the study of solid and hematological tumors.

Acid Nucleic First-generation sequencing technology, second generation and third generation. Methods for the preparation of libraries for the exome analysis and specific regions using a multiplex. Methods of clonal amplification. Techniques for the detection of incorporated nucleotides. Bioinformatics and statistical methods for interpreting data obtained from second-generation sequencing platform (NGS).

Implementation of the first generation sequencing techniques for the detection of mutations with prognostic and predictive significance of response to therapy. Application of second generation technologies for the analysis of solid and hematological tumors.

Detection of chromosomal aberrations by MLPA technology (Multiplex Ligation-dependent Probe Amplification).

Human cell cultures.

Textbook Information

Applied Biochemistry
Fondamenti di Biochimica umana. M. Maccarrone. Zanichelli

Metodologie biochimiche e biomolecolari. M. Maccarrone. Zanichelli
Molecular Diagnostic Technologies
1. Additional material supplied by the teacher; 2) Fundamental Molecular Biology; Lizabeth A. Allison, Editore: John Wiley & Sons Inc; 3) Online courses: http://www.dsf.unict.it/index.php?page=progetto-phar-in; "Riparazione del Dna e mutagenesi nel cancro" (D.F. Condorelli); Mismatch repair (MMR) instabilità dei microsatelliti (V. Barresi); "Tecnologie avanzate per la Bio-Medicina Genomica" (Vincenza Barresi).

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