Overall objectives:

1 - Students should be able to: Acquire basic knowledge about laboratory methods of analysis of nucleic acids and proteins
2 - Know examples of applications of the methodologies to concrete problems and cases of study
3 - Develop practical skills in the use of laboratory equipment and materials, including the use of analytical software
4 - Plan and execute laboratory protocols with autonomy
5 - Recognize risks and apply safety procedures in laboratory practice
6 - Develop teamwork and critical thinking skills
7 - Develop skills in analysis, interpretation and reporting of results
8 - Identify the appropriate methodologies for different studies in the area of Biology
9 - Recognize the limitations and artifacts of each methodology

Syllabus:

1 - Isolation and purification of DNA and RNA
2 - Quantification and quality determination of DNA and RNA (spectrophotometry, Picogreen assays)
3 - Nucleic acid amplification (conventional, nested, multiplex and digital PCR, MLPA, Real Time PCR, RT-qPCR)
4 - Enzyme Restriction Assays (RFLPs)
5 - DNA electrophoresis (agarose, polyacrylamide, DGGE, capillary)
6 - Southern and Northern Blotting
7 - DNA Sequencing (Sanger and NGS)
8 - Genotyping and gene expression microarrays
9 - Molecular cytogenetics (FISH, QF-PCR)
10 - Protein electrophoresis (cellulose acetate, SDS-PAGE, pulse-field, isoelectric focusing, capillary)
11 - Western and Eastern Blotting
12 - Enzymatic methods (spectrophotometry, turbidimetry, electrochemistry, ELISA)
13 - Immunoprecipitation (protein-protein Co-IP, protein-DNA ChIP, protein-RNA RIP / CLIP)
14 - Chromatography (mono and two-dimensional thin layer, ion exchange, gel filtration, affinity, HPLC, gaseous)
15 - Mass spectrometry (MALDI, MALDI-TOF, ESI)

Literature/Sources:

Cseke L.J., Kirakosyan A., Kaufman P.B., Westfall M.V. , 2011 , Handbook of Molecular and Cellular Methods in Biology and Medicine , CRC Press, Taylor & Francis Group
Green M.R., Sambrook J. , 2012 , Molecular Cloning: A Laboratory Manual , Cold Spring Harbor Laboratory Press
Walker J.M., Rapley R. , 2008 , Molecular Biomethods Handbook , Humana Press

Assesssment methods and criteria:

Classification Type: Quantitativa (0-20)

Evaluation Methodology:
The course includes T, TP, PL and S classes, distributed over 14 weeks of instruction and scheduled in order to enable an integrated learning. In T classes the teacher exposes the programmatic contents with the use of audiovisual support. TPs are based on the oriented resolution of exercises and the analysis, discussion and interpretation of raw data, resulting from the molecular methods discussed. PLs include group laboratory work, with application to concrete case studies. The S focus on the discussion of selected scientific articles, which illustrate applications of the methodologies. The course evaluation is based on: - 2 written tests, of integration of T, TP and PL contents, minimum grade 8,0v (30% each) - 2 PL group reports (15% each) - continuous evaluation in the TP and PL (participation, performance, domain of concepts, relevance of interventions, scientific reasoning) (10%).