Content

The course includes 4 types of contents or modules.

Informatics

The practices are organized in 5 sessions of 2 h that take place in the computer rooms.

Practice 1 (2h). Introduction to Bash: First instructions: ls, cd, pwd, ... File manipulation: directory system/folders, redirects, visualization and file sorting, ...

Practice 2 (2h). File manipulation: sort, grep and AWK.

Practice 3 (2h). Introduction to spreadsheets: LibreOffice Calc (I).

Practice 4 (2h). Introduction to spreadsheets: LibreOffice Calc (II).

Practice 5 (2h). Practical application.

Cell Biology

The practices are organized in 6 sessions of 2 h that take place in the laboratory.

Practice 1 (2h). Introduction to the optical microscope and observation of plant cells. Description of the optical microscope elements and instructions for using the microscope. Obtaining temporary preparations from different plant tissue samples (peppers, Elodea) and observing the morphology of plant cells and their main components: cell wall, nucleus, chloroplasts, chromoplasts, plasmodesmata.

Practice 2 (2h). Observation of animal cells under the optical microscope. Observation of the morphology of different types of animal cells: oral mucosa cells, fibroblasts and spermatozoa.

Practice 3 (2h). Introduction to electron microscopy. Fundamentals of electron microscopy. Recognition and measurement of different structures and cell organelles in SEM and TEM micrographs.

Practice 4 (2h). Osmosis and simple diffusion. Study of osmosis in Elodea leaf cells exposed to different concentrations of NaCl. Study of simple diffusion of alcohols across the membrane of Elodea leaf cells.

Practice 5 (2h). The mitotic cell division. Preparation of slides of plant tissues to observe and recognize the different mitotic phasesand estimate their duration.

Practice 6 (2h). The meiotic cell division. Observation of the different phases of the meiotic cycle of spermatogenesis in insects.

Instrumental Techniques

Basic techniques of laboratory work. Application of spectrometry.

Practice 1 (4h). Preparation of a pH buffer system. Determination of glucose concentration using a colorimetric method. Analysis of an absorption spectrum.

Use of the basic techniques of separation, detection and analysis of proteins and nucleic acids.

Practice 2 (4h). Determination of protein concentration using a colorimetric method (Bradford). Separation of proteins by SDS electrophoresis (Part 1). Amplification of a gene by PCR, effect of Mg²⁺ concentration (Part 1).

Practice 3 (4h). Determination of the Mr of some proteins by SDS electrophoresis (example: milk proteins). Separation of DNA fragments by agarose electrophoresis (identification of amplified PCR fragments obtained in practice 2).

Biochemistry

Application of the basic techniques for the analysis, purification and characterization of biomolecules.

Practice 1 (4h). Gel filtration chromatography: separation of hemoglobin from vitamin B12 and blue dextran. Process of expression and purification of a heterologous protein: GFP (green fluorescence protein).

Practice 2 (4h). Continuation of the process of GFP purification. Hydrophobic chromatography: partial purification of GFP from a bacterial extract.

Practice 3 (4h). Identification of lipids by thin layer chromatography. Determination of the pKa of p-nitrophenol and its usefulness to follow the enzymatic activity of phosphatase.

Basic techniques of analysis of enzymatic activity. Study of the activity of acid phosphatase.

Practice 4 (4h). Application of spectrometry to theanalysis of enzymatic activity. Determination of the optimal pH for the activity of an enzyme. Determination of the time in which the linearity of the reaction is maintained. Obtention of initial rate data to determine the kinetic parameters KM and Vm of the reaction. Analysis of the effect of an inhibitor on enzymatic activity.

Practice 5 (2h). Use of computer tools to determine the value of pKa and the kinetic parameters. Use of the GRAFIT program. Determination of pKa of p-nitrophenol from the data obtained in practice 3. Determination of kinetic parameters, KM and Vm, from the data obtained in practice 4. Determination of the type of inhibition and the corresponding inhibition constants from the data obtained in practice 4.