| Activity | Credits | Period | Academic staff | Timetable |
|---|---|---|---|---|
| Teoria | 2 | II semestre | Stefano Capaldi | |
| Laboratorio [II turno] | 4 | II semestre | Matteo Ballottari | |
| Laboratorio [I turno] | 4 | II semestre | Stefano Capaldi |
The course provides an overview of the most common methodologies and techniques for the mani-pulation 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.
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.
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.
| Activity | Author | Title | Publisher | Year | ISBN | Note |
| Teoria | Karcher | LABORATORIO DI BIOLOGIA MOLECOLARE | ed Zanichelli | 1998 | ||
| Teoria | Michael R. Green e Joseph Sambrook | Molecular Cloning: A Laboratory Manual, Fourth Edition (Edizione 4) | CSHL Press | 2012 | 978-1-936113-42-2 | |
| Laboratorio | Karcher | LABORATORIO DI BIOLOGIA MOLECOLARE | ed Zanichelli | 1998 |
