Techniques in molecular and cellular biology

A.Y. 2019/2020
10
Max ECTS
120
Overall hours
SSD
BIO/10 BIO/13
Language
Italian
Learning objectives
The main goal of the course is to introduce and familiarize students with the fundamental laboratory techniques providing a basic understanding of the methodologies employed for the analysis of nucleic acids and proteins. The course will focus on DNA, RNA and proteins purification and characterization, in vivo and in vitro (PCR) molecular cloning. The course will also cover the major principles of enzymatic assays and basic cell culture techniques, including genetic engineering of animal cells. The teaching on the course is split between formal lectures and tutorials most of which are held in dedicated teaching laboratories. This laboratory-based work will provide hands-on experience with nucleic acid and protein isolation, quantification, and characterization. Laboratory activities will include spectroscopy methods (UV-visible), gel electrophoresis (SDS-PAGE, agarose), basic molecular and cell biology techniques such as recombinant DNA, PCR, restriction enzymes, plasmid DNA, sterile techniques, growing bacteria and animal cells, cloning, protein and enzyme assays. Specific goals of the laboratory work is to provide students with the practical skills needed in the basic techniques used in biochemistry, cell and molecular biology, and to let them acquire the knowledge to correctly analyze the experimental results.
Expected learning outcomes
At the completion of this course, students are expected to have an understanding of the major concepts and theoretical principles of basic techniques used in molecular and cell biology as well as the basic practical skills needed to use the fundamental laboratory methodologies.
Course syllabus and organization

Unique edition

Responsible
Lesson period
First semester
Prerequisites for admission
Students must have fulfilled all the prerequisite requirements indicated in the study plan: General and cellular biology, Genetics.
Assessement methods and criteria
The exam consists of a written and oral test
Passing the written test is compulsory for the admission to the oral test
The written test (2 hours) consists of 30 questions (15 for each module) including:
1) multiple choice questions (60%-70 %);
2) short-answer questions (15-20 %);
3) exercises regarding the topics dealt with in practical laboratory classes (15-20 %) (the calculator is allowed).
Each question/exercise achieves 1 point. The test is passed if the score is ≥ 18
The evaluation of a written lab report (optional) on the results obtained in the laboratory classes can increase the score by 1/2 points
The oral consists of 3/4 questions on the course program (lectures and laboratory practice).
Both tests aim to evaluate that the student has theoretical and practical knowledge on the main cellular and molecular methodologies for the separation and analysis of proteins and nucleic acids, and the basic skills to correctly describe and apply the fundamental biochemistry, cellular and molecular biology techniques dealt with in the laboratory experience.
The total course score is the sum of the scores obtained in the written and the oral test (± 4 points). If the exam is not passed, the student will have to take both the written and oral exam.
Cellular methodologies
Course syllabus
Cellular experimental systems: methods in bacterial and animal cell cultures (principles, techniques, and applications).
Cell-based DNA cloning: principles of cell-based molecular cloning, use of restriction endonuclease enzymes, ligation, cloning vectors, transformation, screening of colonies.
Methods for gene cloning and expression: factors affecting gene expression, expression vectors , cellular systems for gene expression (bacteria , yeast, mammalian cells ), markers and signals, mutagenesis.
Gene knockdown methods in cell systems: gene silencing by small interfering RNA (siRNA), microRNA (miRNA), morpholino oligonucleotides.
Teaching methods
The teachers will use: a) Frontal lessons (3 CFU); b): individual and/or small group experimental activity in multidisciplinary laboratories (2CFU).
The teachers will use the Ariel platform to provide the teaching material consisting of: a) copy of the power point presentations used in the lectures; b) handouts with the explanation of the experimental procedures for the laboratory activity; c) video tutorial links of the main techniques and methodologies used in the laboratory experimental activity; d) copy of the technical and safety data sheets of the reagents used in the laboratory experimental activity.
Attendance to lectures and laboratory activities is compulsory.
Bibliography
Dai geni ai genomi -Principi e applicazioni della tecnologia del DNA ricombinante
Jeremy W. Dale, Malcom von Schantz and Nick Plant, Copyright © 2013 EdiSES
PRACTICAL SKILLS IN BIOMOLECULAR SCIENCES
R.Reed, D.Holmes, J.Weyers, A.Jones, Pearson 5th Ed.2016
Molecular methodologies
Course syllabus
Fundamental laboratory techniques: overview and principles of centrifugation, chromatography, electrophoresis and spectrophotometry.
Protein purification: introduction and basic principles; protein quantification methods to determine protein concentration. Analysis of proteins by gel- electrophoresis and blotting.
Methods for determination of enzyme activity: general principles of enzymatic assays.
Extraction, purification and quantification of nucleic acids.
Analysis of nucleic acids by gel-electrophoresis and blotting.
cDNA and genomic libraries, library screening methods.
Molecular cloning using polymerase chain reaction (PCR) technology: principles of the PCR method, optimization, primer design, troubleshooting; cloning of PCR products, types of PCR (Long-Range PCR, RT-PCR, quantitative PCR or 'Real-Time'), applications of PCR.
Analysis of gene expression: Northern blot, RT-PCR, in situ hybridization, reporter genes, promoters, DNA-binding proteins, transcriptome analysis.
Teaching methods
The teachers will use: a) Frontal lessons (2 CFU); b): individual and/or small group experimental activity in multidisciplinary laboratories (3 CFU).
The teachers will use the Ariel platform to provide the teaching material consisting of: a) copy of the power point presentations used in the lectures; b) handouts of the experimental procedures of the laboratory activity; c) video tutorial links of the main techniques and methodologies used in the laboratory experimental activity; d) copy of the technical and safety data sheets of the reagents used in the laboratory experimental activity.
Attendance to lectures and laboratory activities is compulsory.
Bibliography
Dai geni ai genomi -Principi e applicazioni della tecnologia del DNA ricombinante
Jeremy W. Dale, Malcom von Schantz and Nick Plant, Copyright © 2013 EdiSES
PRACTICAL SKILLS IN BIOMOLECULAR SCIENCES
R.Reed, D.Holmes, J.Weyers, A.Jones, Pearson 5th Ed.2016
Cellular methodologies
BIO/13 - EXPERIMENTAL BIOLOGY - University credits: 5
Practicals: 32 hours
Lessons: 24 hours
Molecular methodologies
BIO/10 - BIOCHEMISTRY - University credits: 5
Practicals: 48 hours
Lessons: 16 hours