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CODE 94710
ACADEMIC YEAR 2024/2025
CREDITS
SCIENTIFIC DISCIPLINARY SECTOR BIO/11
LANGUAGE Italian
TEACHING LOCATION
  • GENOVA
SEMESTER 2° Semester
PREREQUISITES
Propedeuticità in ingresso
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TEACHING MATERIALS AULAWEB

OVERVIEW

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 .

AIMS AND CONTENT

LEARNING OUTCOMES

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.

AIMS AND LEARNING OUTCOMES

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.

PREREQUISITES

basic knowledge of biology, physical chemistry and mathematics

TEACHING METHODS

The course consists of 20 lectures and 8 practical laboratory sessions.

SYLLABUS/CONTENT

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"

RECOMMENDED READING/BIBLIOGRAPHY

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

TEACHERS AND EXAM BOARD

LESSONS

LESSONS START

1th oc tober

Class schedule

The timetable for this course is available here: Portale EasyAcademy

EXAMS

EXAM DESCRIPTION

multiple choice written test and oral exam

ASSESSMENT METHODS

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.