The aim of this couse is to provide a description of the basic techniques of recombinant DNA. In particular, the course focuses on the manipulation of nucleic acids, restriction endonuclease digestion, electrophoretic separation of DNA, and cloning. This topics are covered in the theoretical part of the course and in the laboratory as well.
Theoretical part:
Recombinant DNA. Prockaryotic and eukaryotic vectors. Gene cloning. Genomic and cDNA libraries.
Nucleic acid manipulation. Restriction enzyme digestion. Electrophoresis. DNA ligation.
Polymerase Chain Reaction (PCR). Applications.
Bacterial coltures. Trasformation of bacteria and eukaryotic cells.
Recombinant protein production from cloned genes.
Laboratory:
Plasmid DNA purification
Genomic DNA purification
RNA purification
Electrophoresis and DNA purification from gel
Restriction endonuclease digestion
PCR and ligation
Recombinant protein expression and SDS PAGE analysis
Practical course report and oral examination
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