Degree course: 
Corso di First cycle degree in BIOTECHNOLOGY
Academic year when starting the degree: 
Academic year in which the course will be held: 
Course type: 
Supplementary compulsory subjects
Second semester
Standard lectures hours: 
Detail of lecture’s hours: 
Lesson (40 hours), Laboratory (16 hours)

It is recommended to begin the course with a strong background in Cytology/Histology, Molecular Biology and Genetics. Moreover, a good knowledge of English language will be important to read and understand texts and publications that will be provided to the students as teaching material.

Final Examination: 

The learning outcomes will be assessed through an oral interview lasting approximately 25-30 minutes, during which several topics selected from the syllabus will be discussed .The final evaluation will include an overall assessment regarding: 1) the level of knowledge about different topics related to the course content and the precision and overall quality of the answers (50%); 2) the student’s ability to properly motivate his/her statements and to sustain an adequate discussion (30%) and 3) the communication skills (20%). At least one question concerning the laboratory module will be asked. The final mark will be calculated on the above mentioned criteria and the exam will be considered passed equal or over the 18/30 mark.

Voto Finale

The Course of Recombinant DNA technology is aimed at providing the students with the theoretical bases of genetic engineering approaches, coupled to a detailed knowledge of the underlying experimental strategies. This course is organized in two distinct modules, represented by class lessons and a laboratory training section. Class lessons will be focused on a detailed overview of the recombinant DNA methodologies that have been developed in the last four decades, together with a constant reference to the biological problems they were expected to address. Laboratory lessons will provide students the opportunity to autonomously carry out, in a fully equipped molecular biology laboratory, a highly focused experimental plan related to a particular molecular genetics issue. On the basis of the regular and crucial technological support provided by the current recombinant DNA approaches in every facet of modern biotechnologies, this course represents an essential source of knowledge which is in total keeping with the academic purposes of a Bachelor Degree Course in Biotechnology
The expected learning outcomes for this course will be the following:
• Knowledge of the theoretical/operative bases of the main methodologies for DNA isolation, manipulation and analysis.
• Ability to properly develop an experimental plan within the context of the current research in the fields of molecular genetics and molecular biology.
• The ability to carry out bibliography searches and to synthesize the retrieved informations in oral and/or visual representation.
• The ability to achieve an informed judgment, adequate expertise and communication skills in relation to both the experimental approaches and the main scientific achievements in genome sciences.
• The ability to develop both a critic awareness and ability to analyze and discuss issues related to the course contents and the comprehension skills required to develop and maintain issues related to the acquired knowledge, by means of critical reasoning and problem-solving attitudes.

CLASS LESSONS (5 CFUs, 40 hours)

• DNA as an informational molecule
• Molecular cloning: summary
• Second generation cloning strategies
• Second and third generation plasmid-based cloning vectors. Expression vectors
• High-capacity cloning tools: lambda, P1 and cosmid vectors. PAC, Yeast Artificial Chromosomes (YACs) and Bacterial Artificial Chromosomes (BACs). Viral vectors for eucaryotic cells
• Principles of molecular hybridization assays
• Applications of molecular hybridization techniques – 1: Southern, northern and zoo blot. Colony hybridization
• Applications of molecular hybridization techniques – 2: fluorescence labelling of nucleic acids and fluorescence in situ hybridization (FISH), RNA in situ hybridization (RNA ISH), comparative genomic hybridization (CGH), microarray hybridization
• Introduction and applications of PCR: Mutation screening and detection, genomic/cDNA screening, reverse transcription PCR, cloning by PCR, DOP-PCR.
• Real-time and digital PCR : principles and experimental approaches
• Whole genome amplification by PCR: DOP-WGA and Multiple Displacement Amplification (MDA)
• Genomic DNA libraries: introduction and mode of assembly. Library complexity and genome equivalent values. cDNA libraries
• Methods for DNA mutagenesis:
• Gene transfer assays in eucaryotic cells
• Methods for the identification of a gene’s regulatory elements: DNase hypersensitive site’s mapping, gene transfer with reporter vectors, DNA footprinting, Electrophoretic Mobility Shift Assay (EMSA), Chromatin conformation Capture (CCC)
• Introduction to NGS techniques. Overview on RNA-Seq approaches.
• Introduction to the key model organisms in experimental biology. Key concepts for transgenesis. Methods for producing and analyzing transgenic mice
• Transgenic systems with inducible gene expression
• Gene targeting principles and approaches. Gene “knock-out” e “knock-in” in mouse model systems. Conditional gene knock-out
• Chromosome engineering
• Genome editing by means of and zinc-finger nucleases, TALEN and CRISPR/Cas9 assays
• Genetic maps and molecular markers.
• DNA profiling


Four practical laboratory training lessons (4 hours each) will be held. The teaching module is focused on the planning and fulfillment of an experimental plan that wil include the following experiences:

• PCR-mediated analysis of the results of a molecular cloning experiment
• DNA extraction from bacterial clones and analysis by restriction enzymes digestions
• Gene transfer experiments in human cultured cells by means of transient transfection assays
• Validation of the transfection efficacy by realtime PCR gene expression analysis
• Further validation of the transfection efficacy by fluorescence microscopy analysis

Recommended textbook:
1) Watson/Caudy/Myers/Witkowski “DNA ricombinante” – Zanichelli.
2) Maccarone M. “Metodologie biochimiche e biomolecolari” Zanichelli
3) Strachan-Read “Genetica molecolare umana” Zanichelli

Other textbooks or scientific articles might be suggested for in-depth study. The teaching material is updated regularly and will be provided to all students as Powerpoint files, short notes, animation files and articles from scientific literature on selected issues. The abovementioned material will be made available to all students via the e-learning online platform. “Brain training” questions will be uploaded in the online platform. Printed folders describing each exercise to be carried out in the laboratory module will also be provided to each student.


The module will be split in standard class lessons (5 CFU) and an experimental laboratory module (1 CFU). Class lessons will be held with the aid of Power Point slide presentation sessions, coupled to projection of didactic movies when required. A set of “brain training” questions will be regularly uploaded in the e-learning platform in order to give the students the opportunity to self-evaluate their level of knowledge and understanding of the topics explained in the classroom. The experimental laboratory module will be held in the Experimental Biology Lab at the Department of Biotechnology and Life Sciences, via JH Dunant 3, Varese. Each student will be assigned a workstation endowed will all necessary equipment for the implementation of the proposed experimental plan. A printed tutorial guide will also be distributed to the students. Lab attendance is mandatory for all students, who can skip no more than one lesson. Students are required to attend the laboratory lessons with a personal lab coat. Students with known intolerance or allergy to drugs or chemicals that might be present in a research lab must absolutely inform the teacher before the beginning of the lab module.

The teacher is available under appointment for conversations with the students focussed on both the topics discussed in class lessons and organizing issues related to the course, either by phone or e-mail.
Telephone: ++39-0332-421512
Email address: