Unit MOLECULAR AND ANALYTICAL BIOTECHNOLOGIES LABORATORY

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

Hours

• 57 ore - Barbara Camilloni

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 exam includes a written test to be carried out on the LIBREEOL platform, in the presence of the teacher, and at the same time as the written test of the Analytical Chemistry module. The exam consists of a multiple choice test containing both theory questions on the topics covered during the lessons and questions relating to laboratory activities. The test is aimed at ascertaining the level of knowledge pursued by the student and the ability acquired in dealing with application problems similar to those that could arise in a research and/or analysis laboratory. The test grade will be mediated with the grade of the Analytical Chemistry module. The exam is considered passed if the individual module tests have achieved a grade greater than or equal to 18.
Students with disabilities and/or DSA are invited to visit the page dedicated to the tools and measures provided and to agree in advance with the teacher what is necessary (https://www.unipg.it/disabilita-e-dsa).

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

Hours

• 57 ore - Roberta Spaccapelo

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 biology techniques.

Reference texts

Teaching material provided by the professor related to the topics covered.

Educational objectives

The course explores aspects of the biology of microorganisms and the interaction between microorganisms and host. Furthermore, the course aims to provide students with the basic theoretical and practical notions of some of the most important techniques commonly used in the fields of microbiology, molecular biology and biochemistry. A fundamental aspect of the teaching will be to highlight how the different technologies addressed 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

The exam includes a written test to be carried out on the LIBREEOL platform, in the presence of the teacher, and at the same time as the written test of the ANALYTICAL CHEMISTRY LABORATORY module. The exam consists of a multiple choice test containing both theory questions on the topics covered during the lectures and questions relating to laboratory activities. The test is aimed at ascertaining the level of knowledge pursued by the student and the ability acquired in dealing with application problems similar to those that could arise in a research and/or analysis laboratory. The test grade will be averaged with the grade of the ANALYTICAL CHEMISTRY LABORATORY module. The exam is considered passed if the individual module tests have achieved a grade greater than or equal to 18.
Students with disabilities and/or DSA are invited to visit the page dedicated to the tools and measures envisaged and to agree in advance what is necessary with the teacher (https://www.unipg.it/disabilita-e -dsa).

Extended program

1. Microbiology and Virology: Biochemical, genetic, molecular and immunological bases of a limited group of microorganisms necessary to understand the pathogenesis and clinic of the infectious diseases that these organisms transmit. In particular, the following topics will be explored: molecular basis of the microorganism-host interaction; molecular basis of the antigenic variability of viruses, bacteria and protozoa; evasion mechanisms of the immune defenses; cellular and molecular basis of natural immunity against microorganisms; antibiotics: mechanism of action and resistance; structure and replication methods of viruses; in vitro cultivation methods of viruses and quantification of viral suspensions; laboratory methods for the antigenic and genetic characterization of viruses (immunoassays, molecular methods, functional assays).
2. Classical and innovative techniques for the identification of microorganisms: Blood smear and identification of intracellular Plasmodium parasites. Plasmodium cell cycle and disease vectors (mosquitoes). GIEMSA stain and GRAM stain. ELISA test.
3. Biochemistry techniques: determination of the protein concentration of a biological sample; determination of enzymatic units in a biological sample, determination of the kinetic parameters of an enzyme. 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 relating to cellular respiration.
4. Molecular biology techniques: Quantification of DNA and RNA by UV spectrophotometry. Restriction enzyme digestion and restriction map analysis by agarose gel electrophoresis. Preparation of an agarose gel, visualization and analysis of the electrophoretic profile. Setup of a PCR and RT-PCR reaction, vectors and cloning strategies. Southern blot and Northern blot.

Obiettivi Agenda 2030 per lo sviluppo sostenibile

Health and wellness

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
• Alessio Cesaretti

Hours

• 36 ore - Catia Clementi
• 5 ore - Alessio Cesaretti

Language of instruction

Italian

Contents

Principles that underlie analytical chemistry.
Preparation of solutions of known concentration. Determination of the unknown concentration of analytes by constructing a calibration curve

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, pH meter and spectrophotometer.
In particular, the student must acquire the following skills:
• Preparation of solutions of known concentration and their dilution
• 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 lectures), introductory to the laboratory experiences.
There will be also 2-3 laboratory experiences to be held in groups.

Other information

The Professor receives by appointment at her office located in via Pascoli
(ex Casa Rinaldi). To fix an appointment, contact the Professor via email

Learning verification modality

The final exam consists of a written test to be carried out on the LIBREEOL platform in person and together with the written test of the MOLECULAR BIOTECHNOLOGIES LABORATORY module. The test of the ANALYTICAL CHEMISTRY LABORATORY module consists of a multiple choice test containing both theory questions and numerical exercises on the topics covered during the lectures and on the analytical techniques used during the laboratory experiences. The test aims to determine and verify the knowledge level, understanding capability and technical skills achieved by the student on the topics covered during the frontal lectures and the analytical techniques used during the laboratory experiences. It will be tested also the ability of the student in dealing with practical issues similar to those that can be encountered in a chemical research and/or analysis laboratory. The test grade will be averaged with the grade of the MOLECULAR BIOTECHNOLOGY LABORATORY module. The exam is considered passed if the individual module tests have achieved a grade greater than or equal to 18.
Students with disabilities and/or DSA are invited to visit the page dedicated to the tools and measures envisaged and to agree in advance with the teacher what is necessary (https://www.unipg.it/disabilita-e -dsa).

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.
• Stages of an analytical process. Sampling and sample preparation techniques. Definition of evaluation parameters of analytical methods. Response linearity and dinamic range, detection limit and limit of quantification, sensitivity, selectivity, signal to noise ratio, precision, accuracy.
• Measurement uncertainty and its propagation (absolute, relative and relative percentage uncertainty, standard deviation).
• Methods for determining unknown concentrations of analytes; construction of a calibration line. Calibration methods with external standards. Least squares method to derive the equation of the calibration line.
• Applications of spectroscopic techniques, in particular of UV-Visible absorption and emission spectroscopy, to quantitative analysis.

Obiettivi Agenda 2030 per lo sviluppo sostenibile

3 and 4

Canale B

CFU

3

Teacher

Catia Clementi

Teachers

• Catia Clementi
• Alessio Cesaretti

Hours

• 36 ore - Catia Clementi
• 5 ore - Alessio Cesaretti

Language of instruction

Italian

Contents

Principles that underlie analytical chemistry.
Preparation of solutions of known concentration. Determination of the unknown concentration of analytes by constructing a calibration curve

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, pH meter and spectrophotometer.
In particular, the student must acquire the following skills:
• Preparation of solutions of known concentration and their dilution
• 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 lectures), introductory to the laboratory experiences.
There will be also 2-3 laboratory experiences to be held in groups.

Other information

The Professor receives by appointment at her office located in via Pascoli
(ex Casa Rinaldi). To fix an appointment, contact the Professor via email

Learning verification modality

The final exam consists of a written test to be carried out on the LIBREEOL platform in person and together with the written test of the MOLECULAR BIOTECHNOLOGIES LABORATORY module. The test of the ANALYTICAL CHEMISTRY LABORATORY module consists of a multiple choice test containing both theory questions and numerical exercises on the topics covered during the lectures and on the analytical techniques used during the laboratory experiences. The test aims to determine and verify the knowledge level, understanding capability and technical skills achieved by the student on the topics covered during the frontal lectures and the analytical techniques used during the laboratory experiences. It will be tested also the ability of the student in dealing with practical issues similar to those that can be encountered in a chemical research and/or analysis laboratory. The test grade will be averaged with the grade of the MOLECULAR BIOTECHNOLOGY LABORATORY module. The exam is considered passed if the individual module tests have achieved a grade greater than or equal to 18.
Students with disabilities and/or DSA are invited to visit the page dedicated to the tools and measures envisaged and to agree in advance with the teacher what is necessary (https://www.unipg.it/disabilita-e -dsa).

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.
• Stages of an analytical process. Sampling and sample preparation techniques. Definition of evaluation parameters of analytical methods. Response linearity and dinamic range, detection limit and limit of quantification, sensitivity, selectivity, signal to noise ratio, precision, accuracy.
• Measurement uncertainty and its propagation (absolute, relative and relative percentage uncertainty, standard deviation).
• Methods for determining unknown concentrations of analytes; construction of a calibration line. Calibration methods with external standards. Least squares method to derive the equation of the calibration line.
• Applications of spectroscopic techniques, in particular of UV-Visible absorption and emission spectroscopy, to quantitative analysis.

Obiettivi Agenda 2030 per lo sviluppo sostenibile

3 and 4