MOLECULAR DIAGNOSTIC METHODOLOGY
Stampa
Enrollment year
2019/2020
Academic year
2021/2022
Regulations
DM270
Academic discipline
BIO/12 (CLINICAL BIOCHEMISTRY AND BIOLOGY)
Department
DEPARTMENT OF BIOLOGY AND BIOTECHNOLOGY "LAZZARO SPALLANZANI"
Course
BIOTECHNOLOGY
Curriculum
PERCORSO COMUNE
Year of study
Period
2nd semester (01/03/2022 - 14/06/2022)
ECTS
6
Lesson hours
48 lesson hours
Language
Italian
Activity type
WRITTEN TEST
Teacher
PALLADINI GIOVANNI (titolare) - 3 ECTS
NUVOLONE MARIO ULISSE - 3 ECTS
Prerequisites
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Learning outcomes
The student:
1) Will know the main techniques of molecular biology for the extraction, amplification and analysis of nucleic acids
2) Will know the main applications of molecular diagnostics as well as the basic aspects of diagnostic reasoning
Course contents
1. Fundamentals of diagnostic reasoning
2. Nucleic acid extraction techniques
3. Nucleic acid quantification techniques
4. Nucleic acid quality control
5. Nucleic acid amplification techniques: basic principles, main types
6. Polymerase chain reaction: basic principles, assay design, technical considerations, main applications
7. PCR variants: Hot start PCR, Touch down PCR, Nested PCR, Multiplex PCR, Inverse PCR, Direct PCR, GC-rich PCR, Reverse transcriptase polymerase chain reaction (RT-PCR), Quantitative polymerase chain reaction (qPCR), real time qPCR, Reverse transcriptase qPCR (RT-qPCR), digital PCR
8. Nucleic acid electrophoresis
9. Restriction Fragment Length Polymorphism (RFLP)
10. PCR Restriction Fragment Length Polymorphism (PCR-RFLP)
11. High resolution melting analysis
12. Microarrays: SNV chips, gene expression microarrays, Arrays for comparative genomic hybridization (aCGH)
13. In situ hybridization: Fluorescence in situ hybridization (FISH), Spectral karyotyping (SKY)
14. Dideoxy-Termination Sequencing (Sanger Sequencing)
15. Massively parallel sequencing: basic principles
16. Bridge amplification and reversible dye terminator sequencing
17. Real-Time single molecule sequencing with fluorescent nucleotides
18. Target-enrichment strategies for massively parallel sequencing
19. Whole-exome sequencing (WES)
20. Applications of molecular diagnositcs: examples on selected clinical settings
Teaching methods
- Frontal lectures
- Laboratory case discussion
Reccomended or required readings
Tietz textbook of Clinical Chemistry and molecular diagnotics

Ciaccio Lippi - Biochimica Clinica e Medicina di Laboratorio
Assessment methods
Written test (multiple choice test)
Further information