Learning objectives
The aim of the course is to provide an in-depth knowledge of the recombinant DNA technology.
At the end of the course, the student should be able to:
- understand the importance and purpose of gene cloning and learn the main techniques and tools for the manipulation (in vitro and in vivo) of biological macromolecules (KNOWLEDGE AND UNDERSTANDING).
- Design the best strategy to carry out the insertion of a DNA fragment into a cloning vector. Define the analytical methodologies necessary to perform a correct analysis of recombinant clones. Prepare a research report to expose the obtained results (APPLYING KNOWLEDGE AND UNDERSTANDING).
- Know how to evaluate the best methodology to perform a gene cloning experiment (MAKING JUDGEMENTS).
- Expose the results of a gene cloning experiment using proper terminology (COMMUNICATION SKILLS).
- Understand technical and specialist tests in Italian and English (LEARNING SKILLS).
Prerequisites
Some previous knowledge of biology, microbiology, biochemistry and chemistry is highly advisable, as well a good knowledge of English language.
Course unit content
The course will provide in-depth knowledge on the methodologies and strategies underlying recombinant DNA technology.
The following topics will be covered during the course:
- The basics of Gene cloning
- Enzymes used for recombinant DNA technology
- Isolation and purification of genomic and plasmid DNA
- Electrophoresis of nucleic acids
- Cloning vectors for bacteria
- Phage-based vectors: lambda and M13
- In vitro DNA synthesis (PCR)
- Mutagenesis mediated by PCR
- Eukaryotic cloning vectors
- Construction and analysis of genomic libraries
- mRNA purification and cDNA synthesis
- Construction and analysis of cDNA libraries
- DNA sequencing
Full programme
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Bibliography
- Gene Cloning and DNA Analysis: An Introduction, 8th Edition. Terry A. Brown. Zanichelli
- Dai geni ai genomi. Dale, von Schantz – EdiSES
- Analisi dei geni e genomi. Richard J.Reece – EdiSES
- Ingegneria genetica. Principi e tecniche. S. Primrose et al. Zanichelli
- DNA Ricombinante - J.D.Watson et al. Zanichelli
- Molecular biotechnology : principles and applications of recombinant DNA. Glick, Bernard R.
- Molecular biology: principles of genome function. Oxford university press
Teaching methods
The course is organized in:
- theoretical classes by the teacher
- training in laboratory
Assessment methods and criteria
Evaluation of the expected learning achievements will be based on a written test consisting in three open questions on the topics of the course and two questions containing exercises (gene cloning, PCR).
To take the exam, the student must have undergone the laboratory training and prepare a "laboratory report" that will be evaluated during the examination.
The "laboratory report" and the written exam allow to verify the learning ability, independence of judgements, the acquisition of proper terminology, knowledge and application of knowledge to solve practical problems.
Other information
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