66437 - Applied Experimental Biology

Learning outcomes

The student knows the organisms that can be used as model for studying basic mechanisms. The student is able to perform basic activities in cell culture and gene expression experiments. The student is able to plan an experimental design and knows how to process data generated by experimentation.

Course contents

The course is divided into modules:

• Experimental design  
  

1.      Ethical and practical aspects
2.      Control of the variability
3.      Basic statistical tests
4.      Planning the experiment according to the "3RS"
5.      Experimental Unit Definition
6.      Determination of sample size
7.      Identification of appropriate controls
8.      Randomization
9.      Types of Experimental Design
10.      Publication of results

• Evaluation of gene expression by Real Time PCR
  

1.      Primers desig
2.      Choosing the appropriate chemistry
3.      Selection of housekeeping gene
4.      Efficiency analysis
5.      Data processing

•      Molecular  diagnosis
  

1. Genetics of immunoglobulins; definition of lymphoid marker and Ig and TCR gene rearrangements. Heteroduplex analysis and fragment analysis as diagnostic method for detecting Ig and TCR clonal rearrangements. Methods for purifying DNA for the molecular diagnosis.
2. Role of somatic mutations in cancerogenesis. “Drivers” and “passengers” mutations. Techniques used for the highly sensitive detections of somatic mutations in neoplastic tissues.
3. Repeated DNA sequences. Microsatellites as polymorphic markers for parentage testing and pedigree verification.
4.  Techniques for genotyping single nucleotide variations: restriction enzymes, real-time PCR, High resolution melting curve analysis, allele-specific primer extension methods.

Readings/Bibliography

• Festing, M.F.W. et al, The Design of Animal Experiments: Reducing the Use of Animals in Research Through Better Experimental Design. Laboratory Animals Ltd ISBN  1-85315-513-6
• Ruxton G.D.,  Colegrave N.,  Experimental Design for the Life Sciences. Oxford University Press ISBN 978-0-19-928511-2

• http://www.gene-quantification.de/habilitation.html
• http://www.biomart.cn/upload/asset/2008/07/16/1216204807.pdf
• http://www3.appliedbiosystems.com/cms/groups/mcb_support/documents/generaldocuments/cms_040980.pdf

Teaching methods

Preparatory Lectures followed by cooperative learning groups on fundamental topics for theoretical aspects.
Molecular biology lab for practical aspects.

Assessment methods

The principal learning outcomes will be assessed:

-  knowledge of the organisms that can be used as model for biomedical researches

- ability to plan an experiment and to process data

 - knowledge of basic mechanisms of gene expression

- knowledge of basic molecular tools in diagnosis

The final grade will be composed of:

40% on the active participation during lecture and laboratory activities;

60% on oral discussion about lecture topics.

This teaching  is part of an Integrated Course, the final grade will be proportional to the CFU of each teaching:

35106 Policy and Legislation in Animal Biotechnology  3CFU 20%

66437 Applied Experimental Biology 6 CFU 40%

66435 Applied Molecular Biology  6 40%

Teaching tools

Lectures files and material for lerning groups.
Hands on in a fully equipped lab.

Office hours

See the website of Monica Forni

See the website of Augusta Zannoni

See the website of Fabio Gentilini