Methods in Applied Biology

Recommended Prerequisites

Basic knowledge in Biology (Physiology, Ecology, Cell and Molecular Biology) at a BSc level.

Teaching Methods

The course involves theoretical and practical classes and labs.The basic knowledge  and the recent developments of the various methodologies are presented which are then explored in lab classes by performing small projects by the students.

(Examples of work: Expression Analysis of a gene in a cell line, analysis of restriction patterns; particle populations analysis of histograms and cytograms obtained by flow cytometry, the wall forming analysis during pollen tube growth, characterization and identification of animal populations in Ecology).

Learning Outcomes

This course aims to provide students with theoretical and practical knowledge of methodologies and experimental strategies in various areas of biology. Student should identify, analyze and discuss scientific papers in these areas. The student must also acquire technical skills of specimen, cell and molecular manipulation and analysis in order to carry out small projects in the area. Applications range from the cell isolation to the identification and characterization of populations in Ecology.

Work Placement(s)

No

Syllabus

1. Cell Culture: bacterial, plant and animal; methodologies and applications.

2. Microscopy: optical, phase contrast, fluorescence, confocal, multiphoton and electron.

3. Preparation and analysis of biological samples: fixation, inclusion, cryopreservation. Visualization and identification: cytological techniques, immunohistochemistry, immunocytochemistry; Immunohistochemistry and immunofluorescence. Methods of separation and cellular and molecular analysis.

4. Cytometry genome size estimates flow and ploidy level. Cell viability. Introduction to cell ranking.

5. Separation and analysis of proteins: SDS-PAGE electrophoretic techniques; 2-DGE; general coloration and immunodetection. Chromatographic techniques.

6. Separation and analysis of DNA. Electrophoretic techniques, methods of identifying and amplifying DNA sequences (PCR and qPCR), RFLP, etc.

7. Study of the expression of resistance genes to pathogens in plants; identification of bioindicators of soil quality.

Head Lecturer(s)

Paula Cristina Veríssimo Pires

Assessment Methods

Continuous assessment
Laboratory work or Field work: 25.0%
Resolution Problems: 25.0%
Mini Tests: 50.0%

Bibliography

WILSON, K. & WALKER, J. (2010) Principles and Techniques of Biochemistry and Molecular Biology. 7ª ed. Cambridge University Press

SHAPIRO, H. M. (2003) Practical Flow Cytometry. 4ª ed. Wiley-Liss

GIVAN, A. L. (2001) Flow cytometry: First principles. 2ª ed. Wiley-Liss

DOLEZEL, J., GREILHUBER, J. & Suda, J. (2007) Flow Cytometry with Plant Cells: Analysis of Genes, Chromosomes and Genomes. 1ª ed. Wiley-VCH

Buchanan, BB, Gruissem, W., Jones RL (2015) Biochemistry and Molecular Biology of Plants. 2ª ed. Wiley Blackwell.

Murphy, DG, Davidson MW (2013) Fundamentals of light microscopy and electronic imaging. Wiley Blackwell.

Smith, RS (2013) Plant Tissue Culture – Techniques and Experiments. 3ª ed. Academic Press

Glick; B.R. & Pasternack; J.J. (2003). Molecular Biotechnology: Principles and applications of recombinant DNA. ASM Press; Washington; USA

Shena; M. (2002). Microarray analysis. John Wiley & Sons; Inc; New York; USA

Recent papers in the literature (variable from year to year