Biomedical Laboratory techniques

The Molecular Biology course examines aspects related to the structure, organization, replication and repair of the genetic material as well as the flow of genetic information through transcription and translation. Some basic methods for the analysis and amplification of nucleic acids by the Polymerase Chain Reaction are discussed. Detailed program of the course follows. STRUCTURE AND ORGANIZATION OF NUCLEIC ACIDS. Chemical constituents of nucleic acids. Purine and pyrimidine bases, ribose and deoxyribose. Nucleosides and nucleotides. DNA as genetic material (experiments by Griffith, Avery-McLeod-McCarty, Hershey & Chase). Chargaff rules. The double helix of Watson and Crick. Denaturation, renaturation and melting temperature (Tm) of a DNA. RNA: Differences from DNA. Various types of RNA and their biological role: mRNA, rRNA, tRNA. The small RNAs (snRNA, snoRNA). Organization of genetic material: chromatin, nucleosomes, histones and histone modifications. DNA DUPLICATION AND REPAIR. DNA duplication: semiconservative (Meselson & Stahl experiment), bidirectional and semidiscontinuous. DNA polymerases. Replication in E. coli and eukaryotes. The telomere problem: telomerases. DNA damage. Repair systems: photo-repair and excision repair. TRANSCRIPTION. Prokaryotic and eukaryotic RNA polymerases. Transcriptional initiation: prokaryotic and eukaryotic promoters. Elongation and transcriptional termination. The reverse transcriptase. Maturation of eukaryotic mRNAs: capping, polyadenylation, splicing. Outline of transcriptional regulation in prokaryotes and eukaryotes. Prokaryotic operons: the case of the lac operon. Eukaryotic transcription factors. Histone modifications, chromatin remodeling and transcriptional regulation. GENETIC CODE AND TRANSLATION. The genetic code. Mutations. Point mutations, insertions and deletions: the case of hemoglobin S. Expansion of triplets. Translation machinery: mRNA, tRNA, ribosomes and aminoacyl-tRNA synthase. Translation: initiation in prokaryotes and eukaryotes, elongation and termination. TECHNIQUES. Purification and analysis of nucleic acids (electrophoresis, spectrophotometry). Polymerase Chain Reaction: general characteristics and applications. Real Time PCR. DNA fingerprinting.

For an adequate learning, it is recommended that at the beginning of the course the student possesses sufficient knowledge of basic Biology as provided by the Cellular and Molecular Basis of Life course delivered in the first semester of the first year.

The course deals with basic topics which can be found in many textbooks. Those listed below are adequate for exam preparation and allow the student to realize the degree of in-depth study required. - Sadava D, Hillis DM, Heller HC, Hacker S. ELEMENTI DI BIOLOGIA E GENETICA, quinta ediz., Zanichelli 2019. - Raven PH, Johnson GB, Mason KA, et al. Volume 2, GENETICA E BIOLOGIA MOLECOLARE, Piccin 2018.

Lesson attendance is mandatory, at least for 67% of the lectures, and both facilitates the learning process and outlines the knowledge level required at the exam.

Evaluation takes please through a verbal discussion typically consisting of 2-3 questions aimed at assessing the knowledge acquired by the student on the various topics covered during the course. To pass the Molecular Biology module, student must show to have acquired a basic knowledge of structure and functions of nucleic acids and flow of genetic information by transcription and translation. To achieve the highest marks, the student must demonstrate excellent command of the various topics covered during the course and the ability to connect them in a logical and coherent way.

The Molecular Biology course consists in classroom lectures. The different topics covered during the course can be easily found on the recommended textbooks. Auxiliary teaching material may be made available to students on the e-learning website and/or by Classroom. The teacher remains available for clarification on specific topics upon appointment.