The course presents an overview of modern recombinant DNA technologies to provide students with both theoretical and practical fundamental tools of molecular biotechnology, with particular attention to the experimental part and the acquisition of manual skills in the use of a molecular biology laboratory instrumentation.
The course will provide the student with the appropriate knowledge in order to choose, plan and execute the main technologies and techniques of the modern genetic engineering. Due to the significant number of hours of practical training the student has the opportunity to obtain a valuable preparation in the execution of the techniques illustrated in detail during the frontal lessons.
The student will be introduced to the knowledge of:
- Recombinant DNA techniques: historical outline, methodologies
- DNA restriction and ligation enzymes
- Vectors and plasmids
- Cloning in E. coli and applications in eukaryotes
- Gene libraries, molecular probes, DNA marking
- DNA electrophoresis, detection, spectrophotometric assay
- Southern Blot, Northern Blot, applications in DNA fingerprinting
- Hybridization, microarray and RNA interference techniques
- The PCR, design and execution. Identification of possible difficulties and critical points
- Use of PCR in molecular cloning techniques, special cases of PCR
- Sequencing of nucleic acids, principles and main techniques in use
- Production of recombinant proteins; main techniques in use
- Protein interaction studies: protein; The double hybrid in yeast
- Protein interaction studies: protein; main techniques in use
- The GFP as a reporter. Description of the main fields of use
- Main reporter genes and transfection techniques
Basic knowledge of biology, physical chemistry and mathematics.
The course consists of 20 lectures and 8 practical laboratory sessions.
Any Student with documented Specific Learning Disorders (SLD), or with any special needs, should contact the Lecturer(s) and to the dedicated SLD Representative in the Department before class begins, in order to liaise and arrange the specific teaching methods so that the learning aims and outcomes may be met.
Lecture Outline:
Lesson 1: Introduction to recombinant DNA techniques: historical outline, methodologies
Lesson 2: Enzymes of restriction and DNA ligation
Lesson 3: Vectors and plasmids
Lesson 4: Cloning in E. coli and plasmid applications in eukaryotes
Lesson 5: Recombinant proteins in prokaryotes
Lesson 6: Recombinant yeast proteins and problems in the production of recombinant proteins
Lesson 7: Recombinant proteins in animal cells
Lesson 8: gene libraries, molecular probes
Lesson 9: DNA electrophoresis and detection techniques
Lesson 10: southern blot, real time PCR multiplex, micro array
Lesson 11: PCR, design and execution. Identification of possible difficulties and critical points.
Lesson 12: Uses of PCR in molecular cloning techniques, special cases of PCR
Lesson 13: Sequencing of nucleic acids, principles and main techniques in use.
Lesson 14: Protein interaction studies: protein; main techniques in use.
Lesson 15: The GFP as a reporter. Description of the main fields of use.
Lesson 16: Main reporter genes and transfection techniques.
Lesson 17: DNA Profiling techniques and forensic DNA analysis
Lesson 18: DNA Barcoding and its uses
Lesson 19: Parallel Mass Sequencing Methods (NGS)
Lesson 20: DNA analysis in paleogenetics and archaeogenetics
Laboratory Outline:
Lesson 1: PCR
Lesson 2: DNA Precipitation and Restriction Enzyme Digestion
Lesson 3: Electrophoresis and DNA Elution from Agarose Gel
Lesson 4: Ligation and Bacterial Transformation
Lesson 5: Analysis of Transformations and Preparation of Solutions for Plasmid DNA Extraction
Lesson 6: Plasmid DNA Extraction and Restriction Enzyme Digestion
Lesson 7: Electrophoresis and Transfection of Eukaryotic Cells
Lesson 8: Quantitative PCR and Analysis of Results"
Recombinant Technology Notes. Aldo Pagano. Free digital version on the course website
"Molecular biotechnologies", Terry Brown, Ed. Zanichelli; Lecture notes "Recombinant Technology Notes", Aldo Pagano, available on www.biotecnologie.unige.it .; "Recombinant DNA - genes and genomes", Watson JD
Ricevimento: Scheduling an appointment (aldo.pagano@unige.it, Tel: 010-5558213)
Ricevimento: Students may ask for explanations whenever they deem it necessary. They may write an email to soniscarfi@unige.it or by phone: 010 3350227. Scarfì's office it's located in Via A. Pastore 3 ( in front of Mente locale Bar), first floor through the fire escape.
Ricevimento: Scheduling an appointment by e-mail "irene.appolloni@unige.it" or by calling n. 0105558404.
ALDO PAGANO (President)
SONIA SCARFI' (President)
IRENE APPOLLONI
I semester in October.
Multiple choice written test and oral exam.
The written exam will assess the ability to solve genetic engineering problems and design useful reactions. the oral exam will allow to verify the ability to evaluate the appropriate techniques to be used in specific cases. The parameters evaluated are the quality of the exposure, the correct use of the specialized vocabulary, the capacity for critical reasoning on the study carried out.
