STEFANIA MARTUCCIELLO | ANALYTICAL AND BIOMOLECULAR METHODS

cod. 0512800031

ANALYTICAL AND BIOMOLECULAR METHODS

	0512800031
	DEPARTMENT OF CHEMISTRY AND BIOLOGY "ADOLFO ZAMBELLI"
	EQF6
	BIOLOGICAL SCIENCES
	2023/2024

PERCORSO SHARED STUDY PLAN

	OBBLIGATORIO
	YEAR OF COURSE 3
	YEAR OF DIDACTIC SYSTEM 2016
	SPRING SEMESTER

TEACHING UNITS

	SSD	CFU	HOURS	ACTIVITY	TYPE OF ACTIVITY
1	METODOLOGIE BIOMOLECOLARI (MODULO DI METODOLOGIE ANALITICHE E BIOMOLECOLARI)
	BIO/10	5	40	LESSONS	COMPULSORY SUBJECTS, CHARACTERISTIC OF THE CLASS
	BIO/10	1	12	LAB	COMPULSORY SUBJECTS, CHARACTERISTIC OF THE CLASS
2	METODOLOGIE ANALITICHE (MODULO DI METODOLOGIE ANALITICHE E BIOMOLECOLARI)
	CHIM/01	4	32	LESSONS	SUPPLEMENTARY COMPULSORY SUBJECTS
	CHIM/01	2	24	LAB	SUPPLEMENTARY COMPULSORY SUBJECTS

PROFESSORS

	MARIA STRIANESE2 T Curriculum
	STEFANIA MARTUCCIELLO1 Curriculum
	GAETANA PAOLELLA1 Curriculum
	BERNARDINA SCAFURI1 Curriculum

	Objectives
	THE AIM OF THE COURSE IS TO GIVE KNOWLEDGE OF THE BASICS OF THE MAJOR CLASSICAL METHODS OF ANALYSIS AND OF THE MAIN METHODS OF INVESTIGATION IN BIOCHEMISTRY AND MOLECULAR BIOLOGY. THE COURSE ALSO AIMS TO STIMULATE THE DEVELOPMENT OF THE ABILITY TO INTERPRET AND PERFORM AN ANALYTICAL PROTOCOL, AND DESCRIBE THE RESULTS WITH MATHEMATICAL RIGOR, AS WELL AS TO STIMULATE THE DEVELOPMENT OF THE ABILITY TO UNDERSTAND AND APPLY AN EXPERIMENTAL PROTOCOL CONCERNING THE HANDLING OF BIOMOLECULES.

THE AIM OF THE COURSE IS TO GIVE KNOWLEDGE OF THE BASICS OF THE MAJOR CLASSICAL METHODS OF ANALYSIS AND OF THE MAIN METHODS OF INVESTIGATION IN BIOCHEMISTRY AND MOLECULAR BIOLOGY. THE COURSE ALSO AIMS TO STIMULATE THE DEVELOPMENT OF THE ABILITY TO INTERPRET AND PERFORM AN ANALYTICAL PROTOCOL, AND DESCRIBE THE RESULTS WITH MATHEMATICAL RIGOR, AS WELL AS TO STIMULATE THE DEVELOPMENT OF THE ABILITY TO UNDERSTAND AND APPLY AN EXPERIMENTAL PROTOCOL CONCERNING THE HANDLING OF BIOMOLECULES.

	Prerequisites
	ORGANIC CHEMISTRY IS REQUIRED. SKILLS IN MATHEMATICS, PHYSICS, BIOCHEMISTRY AND MOLECULAR BIOLOGY ARE USEFUL.

	Contents
	THE COURSE IS WELL-STRUCTURED INTO TWO MODULES, EACH OF 6 CFU, NAMED MODULE OF ANALYTICAL CHEMISTRY AND MODULE OF BIOMOLECULAR METHODS. EACH MODULE INCLUDES THEORETICAL AND PRACTICAL LECTURES. SUBJECTS OF THE MODULE IF ANALYTICAL CHEMISTRY ARE: INTRODUCTION TO LABORATORY (3 HOURS) DATA ELABORATION AND SELECTION OF ANALYTICAL METHOD (2 HOURS) CHEMICAL EQUILIBRIUM (1 HOUR) ACID-BASE REACTIONS (4 HOURS) ACID-BASE TITRATIONS (3 HOURS) COMPLEXATION REACTIONS (1 HOUR) COMPLEXOMETRIC TITRATION (2 HOURS) INTRODUCTION TO SPECTROSCOPY (2 HOURS) MOLECULAR SPECTROSCOPY (3 HOURS) REDOX REACTIONS (2 HOURS) INTRODUCTION TO ELECTROCHEMISTRY AND POTENTIOMETRY (1 HOUR) REDOX TITRATIONS (3 HOURS) INTRODUCTION TO SEPARATION METHODS (2 HOURS) GASCROMATOGRAPHY (1 HOUR) LIQUID CROMATOGRAPHY (2 HOURS) LAB EXPERIMENTS: 1) ACID-BASE TITRATION. DETERMINATION OF THE CONCENTRATION OF SOLUTIONS OF ACIDS AND BASES BY USING ACID-BASE INDICATORS (4 HOURS). 2) COMPLEXOMETRIC TITRATION. DETERMINATION OF THE WATER HARDNESS BY TITRATION WITH EDTA (4 HOURS). 3-6) IMPLEMENTATION OF A FLUORESCENT BIOSENSOR FOR H2S: I – PREPARATION OF A A BUFFER SOLUTION AND OF THE MYOGLOBIN SAMPLE (4 HOURS). II – UV TITRATION OF MYOGLOBIN WITH H2S (4 HOURS). III – PREPARATION OF FLUORESCENT SAMPLE (4 HOURS). IV – FLUORESCENCE TITRATION OF MYOGLOBIN WITH H2S (4 HOURS). SUBJECTS OF THE MODULE OF BIOMOLECULAR METHODS ARE: CELL CULTURES AND MICROSCOPY. METHODS OF HOMOGENIZATION OF CELLS AND TISSUES (3 HOURS). CENTRIFUGATION (3 HOURS). PROTEINS AND NUCLEIC ACIDS PRECIPITATION (3 HOURS). CHROMATOGRAPHY AND SPECTROPHOTOMETRY APPLIED TO BIOLOGICAL MACROMOLECULES (7 HOURS). ELECTROPHORESIS OF PROTEINS AND NUCLEIC ACIDS (6 HOURS). HYBRIDIZATION AND BLOTTING METHODS. RADIOISOTOPES (4 HOURS). IMMUNOLOGICAL AND IMMUNOENZYMATIC METHODS (4 HOURS). ELEMENTS OF MASS SPECTROMETRY AND PROTEOMICS (3 HOURS). CLONING METHODS AND PCR. INTRODUCTION TO RECOMBINANT DNA TECHNOLOGY (7 HOURS). LABORATORY EXPERIENCES: PRACTICAL EXPERIENCES ARE FOCUSED ON PREPARATION OF PROTEIN EXTRACTS FROM CELL CULTURES, SPECTROPHOTOMETRICAL ANALYSIS, SDS-PAGE AND WESTERN BLOT (12 HOURS).

Contents

THE COURSE IS WELL-STRUCTURED INTO TWO MODULES, EACH OF 6 CFU, NAMED MODULE OF ANALYTICAL CHEMISTRY AND MODULE OF BIOMOLECULAR METHODS. EACH MODULE INCLUDES THEORETICAL AND PRACTICAL LECTURES.

SUBJECTS OF THE MODULE IF ANALYTICAL CHEMISTRY ARE:
INTRODUCTION TO LABORATORY (3 HOURS)
DATA ELABORATION AND SELECTION OF ANALYTICAL METHOD (2 HOURS)
CHEMICAL EQUILIBRIUM (1 HOUR)
ACID-BASE REACTIONS (4 HOURS)
ACID-BASE TITRATIONS (3 HOURS)
COMPLEXATION REACTIONS (1 HOUR)
COMPLEXOMETRIC TITRATION (2 HOURS)
INTRODUCTION TO SPECTROSCOPY (2 HOURS)
MOLECULAR SPECTROSCOPY (3 HOURS)
REDOX REACTIONS (2 HOURS)
INTRODUCTION TO ELECTROCHEMISTRY AND POTENTIOMETRY (1 HOUR)
REDOX TITRATIONS (3 HOURS)
INTRODUCTION TO SEPARATION METHODS (2 HOURS)
GASCROMATOGRAPHY (1 HOUR)
LIQUID CROMATOGRAPHY (2 HOURS)

LAB EXPERIMENTS:
1) ACID-BASE TITRATION. DETERMINATION OF THE CONCENTRATION OF SOLUTIONS OF ACIDS AND BASES BY USING ACID-BASE INDICATORS (4 HOURS).
2) COMPLEXOMETRIC TITRATION. DETERMINATION OF THE WATER HARDNESS BY TITRATION WITH EDTA (4 HOURS).
3-6) IMPLEMENTATION OF A FLUORESCENT BIOSENSOR FOR H2S:
I – PREPARATION OF A A BUFFER SOLUTION AND OF THE MYOGLOBIN SAMPLE (4 HOURS).
II – UV TITRATION OF MYOGLOBIN WITH H2S (4 HOURS).
III – PREPARATION OF FLUORESCENT SAMPLE (4 HOURS).
IV – FLUORESCENCE TITRATION OF MYOGLOBIN WITH H2S (4 HOURS).

SUBJECTS OF THE MODULE OF BIOMOLECULAR METHODS ARE:

CELL CULTURES AND MICROSCOPY. METHODS OF HOMOGENIZATION OF CELLS AND TISSUES (3 HOURS).
CENTRIFUGATION (3 HOURS).
PROTEINS AND NUCLEIC ACIDS PRECIPITATION (3 HOURS).
CHROMATOGRAPHY AND SPECTROPHOTOMETRY APPLIED TO BIOLOGICAL MACROMOLECULES (7 HOURS).
ELECTROPHORESIS OF PROTEINS AND NUCLEIC ACIDS (6 HOURS).
HYBRIDIZATION AND BLOTTING METHODS. RADIOISOTOPES (4 HOURS).
IMMUNOLOGICAL AND IMMUNOENZYMATIC METHODS (4 HOURS). ELEMENTS OF MASS SPECTROMETRY AND PROTEOMICS (3 HOURS).
CLONING METHODS AND PCR. INTRODUCTION TO RECOMBINANT DNA TECHNOLOGY (7 HOURS).

LABORATORY EXPERIENCES:
PRACTICAL EXPERIENCES ARE FOCUSED ON PREPARATION OF PROTEIN EXTRACTS FROM CELL CULTURES, SPECTROPHOTOMETRICAL ANALYSIS, SDS-PAGE AND WESTERN BLOT (12 HOURS).

	Teaching Methods
	LECTURES IN THE CLASSROOM WITH PROJECTION OF TEACHING MATERIALS IN ELECTRONIC FORM. EXERCISES IN CLASS. LABORATORY PRACTISES. 108 HOURS OF LESSONS, EXERCISES AND LABORATORY ARE SPLIT IN TWO MODULES AS FOLLOWS: MODULE: ANALYTICAL CHEMISTRY LESSONS (4 CFU, 32 HOURS). LABORATORY (2 CFU, 24 HOURS). MODULE: BIOMOLECULAR METHODS LESSONS (5 CFU, 40 HOURS). LABORATORY (1 CFU, 12 HOURS). TO ATTEND THEORETICAL LESSONS IS NOT MANDATORY BUT STRONGLY SUGGESTED. TO ATTEND LABORATORY IS MANDATORY. IT IS POSSIBLE TO AFFORD THE EXAM ONLY IF THE STUDENT ATTENDS 70% OF LABORATORY HOURS. THE PRESENCE OF STUDENTS IN LAB IS CONFIRMED BY ROLL CALL MADE BY THE PROFESSOR.

Teaching Methods

LECTURES IN THE CLASSROOM WITH PROJECTION OF TEACHING MATERIALS IN ELECTRONIC FORM. EXERCISES IN CLASS. LABORATORY PRACTISES.

108 HOURS OF LESSONS, EXERCISES AND LABORATORY ARE SPLIT IN TWO MODULES AS FOLLOWS:

MODULE: ANALYTICAL CHEMISTRY
LESSONS (4 CFU, 32 HOURS).
LABORATORY (2 CFU, 24 HOURS).

MODULE: BIOMOLECULAR METHODS
LESSONS (5 CFU, 40 HOURS).
LABORATORY (1 CFU, 12 HOURS).

TO ATTEND THEORETICAL LESSONS IS NOT MANDATORY BUT STRONGLY SUGGESTED.
TO ATTEND LABORATORY IS MANDATORY. IT IS POSSIBLE TO AFFORD THE EXAM ONLY IF THE STUDENT ATTENDS 70% OF LABORATORY HOURS. THE PRESENCE OF STUDENTS IN LAB IS CONFIRMED BY ROLL CALL MADE BY THE PROFESSOR.

	Verification of learning
	THE ACHIEVEMENT OF TEACHING AIMS WILL BE VERIFIED BY MEANS OF A WRITTEN EXAMINATION, INVOLVING BOTH MODULES, WITH A DURATION OF 2 HOUR. THE TEST CONSISTS OF A COMBINATION OF EXERCICES AND QUESTIONS WITH MULTIPLE CHOICES, DIRECT OR OPEN ANSWERS. THE EXAMINATION IS ON THE WHOLE AIMED TO VERIFY THE ACQUISITION OF: KNOWDLEDGE AND EXPERTISES ON METHODS TO STUDY MOLECULES AND MACROMOLECULES; ABILITY TO CORRECTLY APPLY THEORETICAL KNOWLEDGE; ABILITY TO UNDERSTAND PROPOSED PROBLEMS; ADEQUATE LANGUAGE SKILLS.RIGHTS ANSWERS TO QUESTIONS ON BASICAL TOPICS WILL BE CONSIDERED NECESSARY TO PASS THE EXAM. MINIMUM SCORE (18/30) WILL BE ATTRIBUTED TO STUDENTS WITH SUPERFICIAL KNOWLEDGE AND LIMITED COMPREHENSION OF TOPICS AS WELL AS SUFFICIENT CLARITY SKILLS. MAXIMUM SCORE (30/30) WILL BE ATTRIBUTED TO STUDENTS WITH DEEP KNOWLEDGE AND COMPREHENSION OF TOPICS AS WELL AS RIGOROUS CLARITY SKILLS AND GOOD DEDUCTIVE REASONING AND THINKING SKILLS SCORE OF 30/30 CUM LAUDE WILL BE ATTRIBUTED TO STUDENTS WITH EXCELLENT KNOWLEDGE AND COMPREHENSION OF TOPICS AS WELL AS EXCELLENT CLARITY SKILLS AND DEDUCTIVE REASONING AND THINKING SKILLS.

Verification of learning

THE ACHIEVEMENT OF TEACHING AIMS WILL BE VERIFIED BY MEANS OF A WRITTEN EXAMINATION, INVOLVING BOTH MODULES, WITH A DURATION OF 2 HOUR. THE TEST CONSISTS OF A COMBINATION OF EXERCICES AND QUESTIONS WITH MULTIPLE CHOICES, DIRECT OR OPEN ANSWERS.
THE EXAMINATION IS ON THE WHOLE AIMED TO VERIFY THE ACQUISITION OF: KNOWDLEDGE AND EXPERTISES ON METHODS TO STUDY MOLECULES AND MACROMOLECULES; ABILITY TO CORRECTLY APPLY THEORETICAL KNOWLEDGE; ABILITY TO UNDERSTAND PROPOSED PROBLEMS; ADEQUATE LANGUAGE SKILLS.RIGHTS ANSWERS TO QUESTIONS ON BASICAL TOPICS WILL BE CONSIDERED NECESSARY TO PASS THE EXAM.
MINIMUM SCORE (18/30) WILL BE ATTRIBUTED TO STUDENTS WITH SUPERFICIAL KNOWLEDGE AND LIMITED COMPREHENSION OF TOPICS AS WELL AS SUFFICIENT CLARITY SKILLS.
MAXIMUM SCORE (30/30) WILL BE ATTRIBUTED TO STUDENTS WITH DEEP KNOWLEDGE AND COMPREHENSION OF TOPICS AS WELL AS RIGOROUS CLARITY SKILLS AND GOOD DEDUCTIVE REASONING AND THINKING SKILLS
SCORE OF 30/30 CUM LAUDE WILL BE ATTRIBUTED TO STUDENTS WITH EXCELLENT KNOWLEDGE AND COMPREHENSION OF TOPICS AS WELL AS EXCELLENT CLARITY SKILLS AND DEDUCTIVE REASONING AND THINKING SKILLS.

	Texts
	FOR THE MODULE OF ANALYTICAL CHEMISTRY THE FOLLOWING TEXTS ARE ADVISED: D.S. HAGE; J.D. CARR, CHIMICA ANALITICA E ANALISI QUANTITATIVA, PICCIN, PADOVA; D. C. HARRIS, CHIMICA ANALITICA QUANTITATIVA, ZANICHELLI, BOLOGNA. FOR THE MODULE OF BIOMOLECULAR METHODS THE FOLLOWING TEXT IS ADVISED: STOPPINI-BELLOTTI. BIOCHIMICA APPLICATA-EDISES TEXTS FOR CONSULTATION ARE : WILSON, WALKER. BIOCHIMICA E BIOLOGIA MOLECOLARE PRINCIPI E TECNICHE-RAFFAELLO CORTINA EDITORE; REED, HOLMES, WEYERS E JONES, METODOLOGIE DI BASE PER LE SCIENZE BIOMOLECOLARI, ZANICHELLI.

Texts

FOR THE MODULE OF ANALYTICAL CHEMISTRY THE FOLLOWING TEXTS ARE ADVISED:
D.S. HAGE; J.D. CARR, CHIMICA ANALITICA E ANALISI QUANTITATIVA, PICCIN, PADOVA;
D. C. HARRIS, CHIMICA ANALITICA QUANTITATIVA, ZANICHELLI, BOLOGNA.

FOR THE MODULE OF BIOMOLECULAR METHODS THE FOLLOWING TEXT IS ADVISED:
STOPPINI-BELLOTTI. BIOCHIMICA APPLICATA-EDISES
TEXTS FOR CONSULTATION ARE :
WILSON, WALKER. BIOCHIMICA E BIOLOGIA MOLECOLARE PRINCIPI E TECNICHE-RAFFAELLO CORTINA EDITORE;
REED, HOLMES, WEYERS E JONES, METODOLOGIE DI BASE PER LE SCIENZE BIOMOLECOLARI, ZANICHELLI.

	More Information
	FOR THE ACADEMIC YEAR 23/24 PROFESSORS OF THIS TEACHING COURSE ARE: MARIA STRIANESE MSTRIANE@UNISA.IT GAETANA PAOLELLA GPAOLELLA@UNISA.IT

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STEFANIA MARTUCCIELLO | ANALYTICAL AND BIOMOLECULAR METHODS