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ANALYTICAL AND BIOMOLECULAR METHODS

STEFANIA MARTUCCIELLO ANALYTICAL AND BIOMOLECULAR METHODS

0512800031
DEPARTMENT OF CHEMISTRY AND BIOLOGY "ADOLFO ZAMBELLI"
EQF6
BIOLOGICAL SCIENCES
2024/2025

OBBLIGATORIO
YEAR OF COURSE 3
YEAR OF DIDACTIC SYSTEM 2016
FULL ACADEMIC YEAR
CFUHOURSACTIVITY
1METODOLOGIE BIOMOLECOLARI (MODULO DI METODOLOGIE ANALITICHE E BIOMOLECOLARI)
540LESSONS
112LAB
2METODOLOGIE ANALITICHE (MODULO DI METODOLOGIE ANALITICHE E BIOMOLECOLARI)
432LESSONS
224LAB
ANNA MARABOTTI1 T Curriculum
STEFANIA MARTUCCIELLO1 Curriculum
IVANA CAPUTO1 Curriculum
-1
MARIA STRIANESE2 Curriculum
Objectives
GENERAL OBJECTIVE:
THE COURSE AIMS AT PROMOTING KNOWLEDGE OF THE MAIN METHODS OF ANALYSIS AND MAJOR METHODOLOGIES FOR STUDYING BIOLOGICAL MACROMOLECULES

KNOWLEDGE AND UNDERSTANDING
THE STUDENT:
-WILL UNDERSTAND THE PRINCIPLES UNDERLYING METHODOLOGIES FOR THE STUDY OF BIOLOGICAL MOLECULES
-WILL KNOW THE MAIN APPROACHES FOR THE PURIFICATION OF PROTEINS AND NUCLEIC ACIDS FROM BIOLOGICAL SOURCES AND FOR THEIR CHARACTERIZATION
- WILL KNOW HOW TO SELECT AND APPLY THE MAIN METHODS OF ANALYSIS
- WILL UNDERSTAND THE RELATIONSHIPS AMONG THE VARIOUS EXISTING METHODS AND WHAT ARE THE ADVANTAGES OF APPLYING ONE ANALYTICAL METHOD AND NOT ANOTHER
-WILL UNDERSTAND THE POTENTIAL, LIMITATIONS AND COMPLEMENTARITY OF THE DIFFERENT METHODOLOGIES LEARNED

ABILITY TO APPLY KNOWLEDGE AND UNDERSTANDING:
THE STUDENT WILL BE ABLE TO:
- INTERPRET AND EXECUTE ANALYTICAL PROTOCOLS
- EXAMINE AND EXECUTE SIMPLE PROTOCOLS FOR THE STUDY OF BIOLOGICAL MOLECULES
-EXAMINE WITH MATHEMATICAL RIGOR THE RESULTS OF AN ANALYSIS
- APPLY THE MOST IMPORTANT BIOMOLECULAR METHODOLOGIES IN THE STUDY OF RELATIONSHIPS BETWEEN STRUCTURE AND FUNCTION OF BIOLOGICAL MOLECULES
- SOLVE DETERMINATIONS OF UNKNOWN ANALYTES.

AUTONOMY OF JUDGMENT:
THE STUDENT WILL BE ABLE TO:
- EVALUATE THE MOST APPROPRIATE EXPERIMENTAL PROTOCOL RELATED TO THE MANIPULATION OF BIOMOLECULES
- SELECT THE MOST VALID ANALYTICAL PARAMETERS FOR PERFORMING A WELL-DETERMINED ANALYSIS

COMMUNICATION SKILLS:
THE STUDENT WILL BE ABLE TO:
-EXPRESS ACQUIRED KNOWLEDGE IN APPROPRIATE SCIENTIFIC LANGUAGE
- REPRESENT AND COMMUNICATE, USING TEXT OR VIRTUAL TOOLS, RESULTS OF ANALYSES PERFORMED IN THE LABORATORY
- PROCESS OBSERVED RESULTS WITH CRITICALITY.

LEARNING SKILLS:
THE STUDENT WILL BE ABLE TO:
- USE TRADITIONAL BIBLIOGRAPHIC TOOLS AND COMPUTER RESOURCES FOR ANALYSIS AND ARCHIVING;
- PROCEED TO THE CONTINUOUS UPDATING OF THEIR KNOWLEDGE, USING THE TECHNICAL AND SCIENTIFIC LITERATURE.
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, AND INTRODUCTION TO BIOINFORMATICS (12 HOURS).
Teaching Methods
LECTURES IN THE CLASSROOM WITH PROJECTION OF TEACHING MATERIALS IN ELECTRONIC FORM. EXERCISES IN CLASS. LABORATORY PRACTISES.

120 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 EXERCISES 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: "APPLIED BIOCHEMISTRY". EDISES
THE LECTURER WILL INDICATE FROM TIME TO TIME WHICH CHAPTERS AND PARAGRAPHS ARE USEFUL TO CONSULT FOR CONTENT LEARNING

OTHER TEXTS FOR FURTHER STUDY:
BONACCORSI DI PATTI, CONTESTABILE, DI SALVO: "BIOCHEMICAL METHODOLOGIES". AMBROSIAN PUBLISHING HOUSE.
MACCARRONE: "METODOLOGIE BIOCHIMICHE E BIOMOLECOLARI. STRUMENTI E TECNICHE PER IL LABORATORIO DEL NUOVO MILLENNIO", ZANICHELLI
THE LECTURER WILL INDICATE FROM TIME TO TIME WHICH CHAPTERS AND PARAGRAPHS IT IS USEFUL TO CONSULT FOR FURTHER STUDY OF PARTICULAR TOPICS.

OTHER MATERIALS:
AT THE END OF EACH LECTURE THE LECTURER PROVIDES A BULLETED LIST OF ADDITIONAL MATERIALS CONSULTED, ACCESSIBLE ONLINE, PROVIDING THE LINKS DIRECTLY IN THE SLIDES THAT ARE PROVIDED TO STUDENTS.
More Information
THE REGULAR ATTENDANCE AT LECTURES IS STRONGLY RECOMMENDED.
FOR THE ACADEMIC YEAR 24/25 PROFESSORS OF THIS TEACHING COURSE ARE:
MARIA STRIANESE MSTRIANE@UNISA.IT
ANNA MARABOTTI: AMARABOTTI@UNISA.IT
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