96240 - Methods and Techniques in Molecular Biology

Learning outcomes

At the end of the course, the student has a solid theoretical basis for the understanding and the experimental implementation of the main techniques and methodologies used in biomolecular research. In addition, the student develops critical skills for the selection of molecular techniques and methods most suitable for conducting specific scientific investigations, the ability to interpret the data obtained, and evaluating the use of additional alternative or complementary techniques and/or methodologies.

Course contents

The course will explore a range of techniques, methods, and experimental approaches rooted in molecular biology, widely employed across various biological disciplines.

Key techniques to be covered include:

• Purification of nucleic acids and assessment of their concentration, integrity, and purity
• Isolation of DNA and RNA via ultracentrifugation
• Use of DNA probes for the identification and analysis of specific genomic sequences
• Technologies for isolating and manipulating gene and DNA segments (enzimatic digestion, ligation, bacterial transformation) 
• Cloning vectors, prokaryotic and eukaryotic expression vectors, and cosmids
• Expression of heterologous genes in eukaryotic cells (transfection, transduction)
• Polymerase Chain Reaction (PCR)
• DNA sequence determination using chemical methods, Sanger sequencing, and Next-Generation Sequencing (NGS) technologies like Illumina, Nanopore, and SMRT
• Targeted genome editing techniques such as TALEN and CRISPR-Cas9
• Gene expression analysis through methods like Northern Blot, qRT-PCR (SYBR, Taqman probes, digital), microarray, and RNA-sequencing
• Techniques for inhibiting the expression of specific genes, including Antisense RNA, RNA interference, and bacterial non-coding RNA
• Analysis of interactions between biological molecules, including footprinting, EMSA, filter binding, chromatin immunoprecipitation, 3C and its derivatives, and atomic force microscopy
• Determination of RNA secondary and tertiary structure

Readings/Bibliography

Study materials will be provided during the course

Teaching methods

Classroom lessons

Assessment methods

Assessment of learning takes place through the final exam, which ascertains the acquisition of the knowledge and skills expected by carrying out a discussion on the topics covered during the course. The evaluation is out of thirty.

Teaching tools

PC and board

Office hours

See the website of Andrea Vannini