Biomolecular methods

A.Y. 2019/2020
6
Max ECTS
72
Overall hours
SSD
BIO/10
Language
Italian
Learning objectives
The aim of the course is to teach the operational and critical skills necessary for the correct analytical and / or preparative methodology in the field of molecular biotechnology.
Expected learning outcomes
The student will be able to apply techniques and methodologies related to the most frequent experimental problems, such as: separation of cellular organelles, extraction and analysis of proteins from plant material, extraction and analysis of nucleic acids, enzymes purification, design and execution of DNA amplification, gene cloning.
Course syllabus and organization

Single session

Responsible
Lesson period
Second semester
Course syllabus
The program includes the following topics:
- How to work in a laboratory of molecular biotechnology (safety, behavior, equipment, etc.).
- Preparation of solutions (buffer systems, biological buffers, etc.).
- Solubility of proteins and nucleic acids (extraction and precipitation)
- Spectrophotometric techniques (quantification and spectroscopic analysis of nucleic acids and proteins, fluorimetry).
- Electrophoretic techniques: one- and two-dimensional electrophoresis of proteins, electrophoresis of nucleic acids (genomic DNA, restriction fragments, RNA), blotting.
- Immunochemical techniques (Western and dot blotting, ELISA).
- Chromatographic techniques: traditional and MiniSpin (gelfiltration, ion exchange, hydrophobic interaction, affinity).
- Protein and nucleic acids sequencing
- Essays of enzyme activity.
- The enzymes in molecular biology (nucleases, ligases, polymerases, modifying enzymes).
- Restriction enzyme analysis
- PCR: end-point and real-time (design of oligonucleotides, optimum temperature, titration of MgCl2).
- Ligation of a restriction fragment in a plasmid cloning. Transformation and blue-white screen.
- Bioinformatics: bioinformatics in gene cloning, PCR and protein studies (databases, search of sequences for alignment, position in the genome, etc.).

[Program for not attending students with reference to descriptor 1 and 2]:
The program includes the following topics:
- How to work in a laboratory of molecular biotechnology (safety, behavior, equipment, etc.).
- Preparation of solutions (buffer systems, biological buffers, etc.).
- Solubility of proteins and nucleic acids (extraction and precipitation)
- Spectrophotometric techniques (quantification and spectroscopic analysis of nucleic acids and proteins, fluorimetry).
- Electrophoretic techniques: one- and two-dimensional electrophoresis of proteins, electrophoresis of nucleic acids (genomic DNA, restriction fragments, RNA), blotting.
- Immunochemical techniques (Western and dot blotting, ELISA).
- Chromatographic techniques: traditional and MiniSpin (gelfiltration, ion exchange, hydrophobic interaction, affinity).
- Protein and nucleic acids sequencing
- Essays of enzyme activity.
- The enzymes in molecular biology (nucleases, ligases, polymerases, modifying enzymes).
- Restriction enzyme analysis
- PCR: end-point and real-time (design of oligonucleotides, optimum temperature, titration of MgCl2).
- Ligation of a restriction fragment in a plasmid cloning. Transformation and blue-white screen.
- Bioinformatics: bioinformatics in gene cloning, PCR and protein studies (databases, search of sequences for alignment, position in the genome, etc.).
Prerequisites for admission
Biochemistry principles of biological macromolecules in particular proteins and nucleic acids.
Teaching methods
Fronal lessons and pratical classes
Teaching Resources
ARIEL web site
Assessment methods and Criteria
The exam consists of a written test lasting 90 minutes that includes both theoretical questions and numerical exercises related to the program carried out during the course.
The exam can be taken only after having attended the practical exercises and after the presentation of the "Laboratory notebook" relating to the experiences carried out. It is mandatoty to sign up at the exam within the deadline indicated on the SIFA. Evaluation is expressed in thirtieths. The final grade will also include the evaluation of the laboratory notebook. Results are communicated to the students by an automatically-generated e-mail.
BIO/10 - BIOCHEMISTRY - University credits: 6
Single bench laboratory practical: 48 hours
Lessons: 24 hours
Professor: Magni Chiara
Professor(s)
Reception:
Contact the professor
DeFens, sez. Scienze Chimiche e Biomolecolari