Unit MOLECULAR AND ANALYTICAL BIOTECHNOLOGIES LABORATORY

Course
Biotechnology
Study-unit Code
A000976
Curriculum
In all curricula
CFU
9
Course Regulation
Coorte 2021
Offered
2022/23
Type of study-unit
Obbligatorio (Required)
Type of learning activities
Attività formativa integrata

MOLECULAR BIOTECHNOLOGY LABORATORY

Code 55072403
CFU 6
Learning activities Caratterizzante
Area Discipline biotecnologiche con finalità specifiche:mediche e terapeutiche
Academic discipline MED/07
Type of study-unit Obbligatorio (Required)

Canale A

CFU
6
Teacher
Barbara Camilloni
Teachers
  • Barbara Camilloni
  • Manlio Di Cristina
  • Brunella Tancini
Hours
  • 57 ore - Barbara Camilloni
  • 10 ore - Manlio Di Cristina
  • 5 ore - Brunella Tancini
Language of instruction
Italian
Contents
Microbiology and Virology: biochemical, genetic, molecular and immunological bases of microorganisms necessary for understanding the pathogenesis and clinical of the infectious diseases that these organisms transmit; Next generation sequencing techniques (NGS) for the identification of microorganisms; Biochemistry techniques; Molecular Genetics techniques; Molecular Biology Techniques; Cytofluorimetry techniques.
Reference texts
Teaching material provided by the Professor related to the topics covered.
Educational objectives
The course aims to provide students with the theoretical and practical basics of some of the most important techniques commonly used in molecular biotechnology. To this end, microbiology, biochemistry, molecular genetics and molecular biology techniques will be discussed. Sequencing techniques will also be analyzed with particular attention to the most innovative new generation techniques (next-generation sequencing - NGS). A fundamental aspect of teaching will be to highlight how the different technologies discussed can be integrated to answer complex biological questions.
Prerequisites
No prerequisites required.
Teaching methods
The course will be held through lectures in which the professor explains the basics, also through the aid of visual aids, and provides practical examples to facilitate their understanding, through practical exercises in the laboratory related to the techniques explained and the use of interactive laboratories.
Other information
Attendance is not mandatory, but strongly recommended.
Lessons will be held at the Biotechnology Pole in Via del Giochetto, Monteluce area.
Learning verification modality
The student will have to take an oral exam aimed at verifying: the ability to understand the main aspects of the topics covered during the course; the ability to develop independently a specific experimental design and to evaluate critically the results obtained through the application of the same techniques; the quality of the oral exposure
Extended program
1. Microbiology and Virology: Biochemical, genetic, molecular and immunological bases of a limited group of microorganisms necessary for understanding the pathogenesis and clinical of the infectious diseases that these organisms transmit. In particular, the following topics will be explored: molecular bases of the microorganism-host interaction; molecular bases of the antigenic variability of viruses and bacteria; mechanisms of evasion of immune defenses; cellular and molecular bases of natural immunity against microorganisms; antibiotics: mechanism of action and resistance; structure and replication of viruses; methods of in vitro cultivation of viruses and quantification of viral suspensions; laboratory methods for the antigenic and genetic characterization of viruses (immunometric assays, molecular methods, functional assays).
2. New generation sequencing techniques (next generation sequencing - NGS) for the identification of microorganisms. Analysis of protocols for library preparation for NGS and bioinformatics pipeline. Virtual laboratory: it involves the active participation of the student through a virtual path that will guide him in the design and implementation of an experiment concerning gene expression.
3. Biochemistry techniques: determination of the protein concentration of a biological sample; determination of the enzymatic units in a biological sample, determination of the kinetic parameters of an enzyme. Virtual laboratory: it involves the active participation of the student through a virtual path that will guide him in the design and implementation of an experiment related to cellular respiration.
4. Molecular Genetics Techniques: Extraction of genomic DNA from Swab. PCR-RFLP analysis techniques and visualization of the restriction profile by agarose gel electrophoresis. Study of genomic variability by bioinformatics programs and phylogenetic analysis. Virtual laboratory: it involves the active participation of the student through a virtual path that will guide him in the design and implementation of an experiment pertaining to forensic genetics.
5. Molecular Biology Techniques: Extraction of DNA from plasmid vectors. DNA quantification using UV spectrophotometry. Digestion with restriction enzymes and analysis of the restriction map by agarose gel electrophoresis. Virtual laboratory: path concerning the design and implementation of a cloning experiment.
6. Cytofluorimetry techniques: Cytofluorimetric analysis of surface antigens in cellular, normal and tumor subpopulations. Cytofluorimetric analysis of intracellular antigens in cellular, normal and tumor subpopulations. Cytofluorimetric analysis of the cycle of cellular, normal and tumoral populations.

Canale B

CFU
6
Teacher
Roberta Spaccapelo
Teachers
  • Roberta Spaccapelo
  • Manlio Di Cristina
  • Brunella Tancini
Hours
  • 57 ore - Roberta Spaccapelo
  • 10 ore - Manlio Di Cristina
  • 5 ore - Brunella Tancini
Language of instruction
ITALIAN
Contents
The course is divided into interactive lectures and laboratory activities. The lectures will deal with Microbiology and Virology topics aimed at understanding the biochemical, genetic, molecular and immunological bases of the microorganisms necessary to explain the pathogenesis of infectious diseases. Practical laboratory activities include: classic and innovative techniques for the identification of microorganisms; Biochemistry techniques; Molecular genetics techniques; Molecular biology techniques; Cytofluorimetry techniques.
Reference texts

Teaching material provided by the professor related to the topics covered.
Educational objectives
The course aims to provide students with the theoretical and practical basics of some of the most important techniques commonly used in molecular biotechnology. To this end, microbiology, biochemistry, molecular genetics, molecular biology and cell therapies techniques will be discussed. Sequencing techniques will also be analyzed with particular attention to the most innovative new generation techniques (next-generation sequencing - NGS). A fundamental aspect of teaching will be to highlight how the different technologies discussed can be integrated to answer complex biological questions.
Prerequisites

To take the exam it is necessary to have passed and registered the preliminary exam of GENERAL MICROBIOLOGY.
Teaching methods
The course will take place through lectures in which the teacher exposes the basics, also through the aid of visual aids, and provides practical examples to facilitate their understanding, through laboratory exercises relating to the techniques addressed and the use of interactive laboratories.
Other information

Attendance is not mandatory, but strongly recommended.
Lessons will be held at the Biotechnology Pole in Via del Giochetto, Monteluce area.
Learning verification modality
Written examination aimed at verifying the ability to understand the main aspects concerning the topics dealt with during the lectures and the ability to understand and carry out the practical activities carried out during the workshops of the course.
Extended program
1. Microbiology and Virology: Biochemical, genetic, molecular and immunological bases of a limited group of microorganisms necessary for understanding the pathogenesis of the infectious diseases that these organisms transmit. In particular, the following topics will be explored: molecular bases of the microorganism-host interaction; molecular bases of the antigenic variability of viruses, bacteria and protozoa; mechanisms of evasion of immune defenses; cellular and molecular bases of natural immunity against microorganisms; antibiotics: mechanism of action and resistance; structure and replication of viruses; methods of in vitro cultivation of viruses and quantification of viral suspensions; laboratory methods for the antigenic and genetic characterization of viruses (immunometric assays, molecular methods, functional assays).

2. Classic and innovative techniques for the identification of microorganisms: staining, ELISA test and Next generation sequencing (NGS). Virtual laboratory: provides for the active participation of the student through a virtual path that will guide him in the design and implementation of an experiment related to gene expression, GRAM staining, ELISA and NGS tests.

3. Biochemistry techniques: determination of the protein concentration of a biological sample; determination of the enzymatic units in a biological sample, determination of the kinetic parameters of an enzyme. Virtual laboratory: it involves the active participation of the student through a virtual path that will guide him in the design and implementation of an experiment related to cellular respiration.
4. Molecular Genetics Techniques: Preparation of a PCR reaction. Control of the reaction product by Electrophoretic Gel. Data interpretation using qualitative and quantitative markers. Genomic DNA extraction starting from saliva. Preparation of PCR reactions, electrophoretic travel on agarose gel, visualization and analysis of the electrophoretic profile. Virtual laboratory: PCR
5. Molecular Biology Techniques: Extraction of DNA from plasmid vectors. DNA quantification using UV spectrophotometry. Digestion with restriction enzymes and analysis of the restriction map by agarose gel electrophoresis. Virtual laboratory: path concerning the design and implementation of a cloning experiment.
6. Cytofluorimetry techniques: Cytofluorimetric analysis of surface antigens in cellular, normal and tumor subpopulations. Cytofluorimetric analysis of intracellular antigens in cellular, normal and tumor subpopulations. Cytofluorimetric analysis of the cycle of cellular, normal and tumoral populations. Virtual laboratory: FACS analysis

ANALYTICAL CHEMISTRY LABORATORY

Code M0017
CFU 3
Learning activities Affine/integrativa
Area Attività formative affini o integrative
Academic discipline CHIM/01
Type of study-unit Obbligatorio (Required)

Canale A

CFU
3
Teacher
Catia Clementi
Teachers
  • Catia Clementi
Hours
  • 36 ore - Catia Clementi
Language of instruction
Italian
Contents

Reference texts
• Teaching materials.

• Lecture notes concerning laboratory experiences

D.C. Harris, “Chimica Analitica Quantitativa”, Zanichelli, Bologna,

F. W. Fifield, D. Kealey, “Chimica Analitica teoria e pratica”, Zanichelli, Bologna

Skoog,. West, Holler, Crouch, “Fondamenti di Chimica Analitica”, EdiSES, Napoli
Educational objectives
The main aim of the course is the understanding of the principles that underlie analytical chemistry and the learning of techniques and methodologies for the preparation and analysis of samples that may be useful in the activity of a biochemical-clinical laboratory. Learn how to manage basic equipment of an analytical chemistry laboratory, such as glassware, burettes, automatic pipettes, beakers, flasks, scales and spectrophotometer.
In particular, the student must acquire the following skills:
• Preparation of solutions of known concentration by weighing and dilution and determination of the relative uncertainties
• Preparation of solutions of known pH
• Determination of the unknown concentration of analytes by constructing a calibration curve
• Use of UV-Visible spectroscopy in absorption and emission for the determination of analytes in unknown samples.
Prerequisites
For a full and easy understanding of the topics of this course it is important to have good basic knowledge of General Chemistry and Physical Chemistry.
Teaching methods
The course includes some theoretical lessons (about 10/15 hours of frontal lectures). Laboratory and seminar activities according to the number of students and the available spaces.

.
Other information

Learning verification modality
The final exam consists of a written test lasting about 30 minutes containing exercises and multiple choice theoretical questions relating to the topics covered in the theoretical and laboratory lessons.
Extended program
• Definition of the basic concepts and purposes of analytical chemistry. Basic tools of the analytical laboratory. Classification of analytical methods: qualitative and quantitative analysis, classical and instrumental methods of analysis, absolute and comparative methods.
• Uncertainties and significant figures in measurements. Concepts of precision and accuracy, absolute and random error: determination and propagation.
• Preparation of solutions with known concentration by weighing and dilution and determination of their relative uncertainties.
• Stages of an analytical process. Sampling and sample preparation techniques. Definition of evaluation parameters of analytical methods. Response linearity range, detection limit and dosability, sensitivity, selectivity, signal to noise ratio, precision, accuracy.
• Methods for determining unknown concentrations of analytes; construction of a calibration line. Calibration methods with external standards, with internal standards and standard addition method.
• Applications of spectroscopic techniques, in particular of UV-Visible absorption and emission spectroscopy, to quantitative analysis.

Canale B

CFU
3
Teacher
Catia Clementi
Teachers
  • Catia Clementi
Hours
  • 36 ore - Catia Clementi
Language of instruction
Italian
Contents

Reference texts
• Teaching materials.

• Lecture notes concerning laboratory experiences

D.C. Harris, “Chimica Analitica Quantitativa”, Zanichelli, Bologna,

F. W. Fifield, D. Kealey, “Chimica Analitica teoria e pratica”, Zanichelli, Bologna

Skoog,. West, Holler, Crouch, “Fondamenti di Chimica Analitica”, EdiSES, Napoli
Educational objectives
The main aim of the course is the understanding of the principles that underlie analytical chemistry and the learning of techniques and methodologies for the preparation and analysis of samples that may be useful in the activity of a biochemical-clinical laboratory. Learn how to manage basic equipment of an analytical chemistry laboratory, such as glassware, burettes, automatic pipettes, beakers, flasks, scales and spectrophotometer.
In particular, the student must acquire the following skills:
• Preparation of solutions of known concentration by weighing and dilution and determination of the relative uncertainties
• Preparation of solutions of known pH
• Determination of the unknown concentration of analytes by constructing a calibration curve
• Use of UV-Visible spectroscopy in absorption and emission for the determination of analytes in unknown samples.
Prerequisites
For a full and easy understanding of the topics of this course it is important to have good basic knowledge of General Chemistry and Physical Chemistry.
Teaching methods
The course includes some theoretical lessons (about 10/15 hours of frontal lectures). Laboratory and seminar activities according to the number of students and the available spaces.

.
Other information

Learning verification modality
The final exam consists of a written test lasting about 30 minutes containing exercises and multiple choice theoretical questions relating to the topics covered in the theoretical and laboratory lessons.
Extended program
• Definition of the basic concepts and purposes of analytical chemistry. Basic tools of the analytical laboratory. Classification of analytical methods: qualitative and quantitative analysis, classical and instrumental methods of analysis, absolute and comparative methods.
• Uncertainties and significant figures in measurements. Concepts of precision and accuracy, absolute and random error: determination and propagation.
• Preparation of solutions with known concentration by weighing and dilution and determination of their relative uncertainties.
• Stages of an analytical process. Sampling and sample preparation techniques. Definition of evaluation parameters of analytical methods. Response linearity range, detection limit and dosability, sensitivity, selectivity, signal to noise ratio, precision, accuracy.
• Methods for determining unknown concentrations of analytes; construction of a calibration line. Calibration methods with external standards, with internal standards and standard addition method.
• Applications of spectroscopic techniques, in particular of UV-Visible absorption and emission spectroscopy, to quantitative analysis.
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