Molecular biology workshop (2019/2020)



Course code
4S01911
Credits
6
Coordinator
Stefano Capaldi
Academic sector
BIO/11 - MOLECULAR BIOLOGY
Language of instruction
Italian
Teaching is organised as follows:
Activity Credits Period Academic staff Timetable
Teoria 2 II semestre Stefano Capaldi

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Laboratorio [II turno] 4 II semestre Matteo Ballottari

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Laboratorio [I turno] 4 II semestre Stefano Capaldi

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Learning outcomes

The course provides an overview of the most common methodologies and techniques for the ma-nipulation of DNA. In particular, the students will learn the main techniques for purification of nucleic acids, their separation by electrophoresis, DNA amplification and methods for cloning in bacterial vectors. In the practical exercises the student will apply these techniques for purification of plasmid DNA and genomic DNA, separation of DNA by electrophoresis on agarose gel, its digestion with re-striction enzymes, preparation of gene constructs and their transformation into E. coli. At the end of the course, the student will have learned the basics of manipulation of genetic material and will be able to use the main techniques for gene cloning.

Syllabus

Theory:
The recombinant DNA. Cloning of Genes.
Bacterial and non bacterial vectors. cDNA and genomic libraries.
Manipulation of nucleic acids. Digestion with restriction endonucleases. Electrophoresis. Ligation of the DNA.
Polymerase Chain Reaction (PCR) and its applications.
Bacteria colutres. Trasformation of bacterial and eukaryotic cells.
Production of recombinant proteins.

Laboratory experiences:
Purification of plasmid DNA
Purification of genomic DNA
Purification of RNA
Electrophoresis and purification of DNA from gel.
Digestion with restriction enzymes.
PCR and PCR product purification.
Transformation, colony PCR
Expression of a recombinant protein and its analysis by SDS PAGE and Western Blot.

Assessment methods and criteria

The final exam consists of a short report on the laboratory activity (to be delivered before the exam session) and an oral interview. The vote will be the average of the two tests.

Reference books
Activity Author Title Publisher Year ISBN Note
Teoria Jeremy W. Dale e Malcolm von Schantz Dai geni ai genomi (Edizione 2) EdiSES 2012
Teoria Karcher LABORATORIO DI BIOLOGIA MOLECOLARE ed Zanichelli 1998
Laboratorio Jeremy W. Dale e Malcolm von Schantz Dai geni ai genomi (Edizione 2) EdiSES 2012
Laboratorio Karcher LABORATORIO DI BIOLOGIA MOLECOLARE ed Zanichelli 1998