Learning objectives
The aim of the course is to provide a theoretical knowledge and practical application of the recombinant technologies.
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 even to a non-expert public (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
- Fundamental principles 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 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, 7th 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, followed by the individual work by
the student for preparation;
- training in laboratory.
Assessment methods and criteria
Students can choose between two ways of examination:
- a written test, consisting in three open questions on the topics of the course and two questions containing exercises (gene cloning, PCR).
- the oral examination on the whole programme (Erasmus students; italian students who have taken the course before 2013/2014) .
In both cases to be admitted to the oral examination the student should prepare a report on laboratory training that will be evaluated during the examination.
The written exam and the "laboratory report" verifies knowledge and application of knowledge to solve practical problems. The oral examination verifies the knowledge acquired by the student and the communication skills. In all cases, the learning ability is verified.
Other information
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