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RECOMBINANT DNA TECHNOLOGIES AND LABORATORY

CODE 94710
ACADEMIC YEAR 2022/2023
CREDITS
  • 7 cfu during the 2nd year of 8756 BIOTECNOLOGIE (L-2) - GENOVA
  • SCIENTIFIC DISCIPLINARY SECTOR BIO/11
    LANGUAGE Italian
    TEACHING LOCATION
  • GENOVA
  • SEMESTER 2° Semester
    PREREQUISITES
    Prerequisites
    You can take the exam for this unit if you passed the following exam(s):
    • Biotechnology 8756 (coorte 2021/2022)
    • BIOLOGY AND GENETICS 80754
    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 laboratory exercises.

    SYLLABUS/CONTENT

    Content program

    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

    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

    Exam Board

    SONIA SCARFI' (President)

    ALDO PAGANO (President)

    IRENE APPOLLONI

    LESSONS

    LESSONS START

    1th oc tober

    Class schedule

    All class schedules are posted on the EasyAcademy portal.

    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.