Laboratory Techniques in Biology II
Code
8143
Academic unit
Faculdade de Ciências e Tecnologia
Department
Departamento de Ciências da Vida
Credits
6.0
Teacher in charge
Ana Madalena Peres de Drumond Ludovice Mendes Gomes, Rita Gonçalves Sobral Almeida
Weekly hours
5
Total hours
68
Teaching language
Português
Objectives
At the end of the semester it is expected that students have acquired the following skills: (1) Describe and explain the molecular basis associated with the several techniques discussed during the course. (2) Distinguish and describe the molecular biology methods that are used to isolate, purify and characterize nucleic acids (gel electrophoresis (agarose), restriction mapping, PCR, in situ hybridization of nucleic acids and immunocytochemistry). (3) Compare and contrast the different approaches described throughout the course. Critically associate these differences to the desired end goal. (4) Regarding a specific biological question, identify the experimental approach most appropriate to address a specific problem. (5) Analyze and interpret experimental results, critically and clearly. Present the results in the form of a scientific report. (6) Understand the importance of technical evolution over the years and that the methodologies are technically limited to the existing knowledge. (7) Understand that molecular and cellular biology are areas in constantly development, always on the edge of the latest technical breakthroughs. (8) Acquire skills to subsequently attend Molecular Genetics and Genetic Engineering Course Units.
Prerequisites
Attendance to Laboratory Techniques I and Cell Biology is highly advised and recommended.
Subject matter
Introduction to the Molecular Biology Laboratory. Overview of risks and safety procedures.
1. Molecular Biology Techniques:
- methods for nucleic acid extraction and purification
- quantitative and qualitative analysis of DNA samples;
- DNA electrophoresis (types and applications);
- introduction to DNA sequencing methods;
- restriction enzyme digestion and restriction maps for genetic analysis;
- PCR analysis – relevance of primer design and experimental conditions.
- bacterial transformation with plasmid DNA molecules.
2. Cyto-molecular techniques:
- fixation and sample preparation for histological analysis;
- histological staining techniques;
- preparation of microscopy slides;
- DNA:DNA fluorescent in situ hybridization.
- Introduction to epifluorescence microscopy
Case-studies of molecular biology applied to genetic analysis.
Bibliography
- Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P. Molecular Biology of the Cell. 6th ed. Garland Science, New York, USA, 2008.
- Reed, R., Holmes, D., Weyers, J., Jones, A. Practical Skills in Biomolecular Sciences. 2nd ed. Pearson Education Limited, Essex, UK, 2003.
- Cantor, C.R. and Smith, C.L. Genomics: The Science and Technology Behind the Human Genome Project. John Wiley & Sons, 1999.
- Sambrook J., and D. W. Russell. (2001) Molecular Cloning - A Laboratory Manual. 3rd Edition. Cold Spring Harbor Laboratory Press. New York.
- Class handouts and protocols for each lab session.
- Research articles.
Teaching method
Theoretical analysis of techniques, methodologies and experimental designs will be covered by the theoretical-practical lectures.
Practical course in laboratory, with fundamental experiments allowing the direct use of molecular and cyto-molecular biology techniques; analysis of results.
Evaluation method
In order to have access to evaluation, the student must attend 2/3 of the total practical classes, corresponding to 8 of the 13 planned classes.
The final grade of the course is obtained through the following criteria:
- A written report on the first experimental work, performed by each group. The assessment of this report contributes with 15% to the final grade (3/20 points).
- Two mini-tests to be carried out during TP classes, each contributing with 10% to the final grade (2/20 points each mini-test).
- The laboratory performance during the semester will contribute with 5% to the final grade (1/20 points).
- Two partial tests or an Exam, being under assessment the theoretical and practical contents, will contribute with 60% to the final grade (12/20 points).