Laboratory Techniques in Biology II
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, hybridization of nucleic acids and sequencing). (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.
Ana Madalena Peres de Drumond Ludovice Mendes Gomes, Rita Gonçalves Sobral de Almeida
Weekly - 5
Total - 68
Attendance to Laboratory Techniques I and Cell Biology is highly advised and recommended.
- Brown, T.A. Gene cloning and DNA analysis - an introduction. 6th ed. Wiley-Blackwell. 2010.
- 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.
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.
In order to have access to evaluation, the student must attend 11 of the total practical classes and 11 of the theoretical-practical classes.
The final grade of the course is obtained through the following criteria:
- Three partial tests or an Exam, being under assessment the theoretical and practical contents, will contribute with 75% to the final grade (15/20 points).
- A written report on the first experimental work, performed by each group. The assessment of this report contributes with 17,5% to the final grade (3,5/20 points).
- Experimental worksheets, performed by each group, and delivered during the corresponding practical lesson, will contribute with 7,5% to the final grade (1,5/20 points).
- The average of the three partial tests must be equal or higher than 9,5
Introduction to the Molecular Biology Laboratory. Overview of risks and safety procedures.
Molecular Biology Techniques
- methods for nucleic acid extraction and purification
- quantitative and qualitative analysis of DNA samples;
- DNA electrophoresis (types and applications);
- restriction enzyme digestion and restriction maps for genetic analysis;
- bacterial transformation with plasmid DNA molecules;
- PCR analysis – relevance of primer design and experimental conditions;
- introduction to DNA sequencing methods;
- genome sequencing;
- transcript analysis;
- protein analysis
Programs where the course is taught: