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Applied Molecular Biology

Code 8435
Year 1
Semester S1
ECTS Credits 6
Workload TP(60H)
Scientific area Biochemistry
Entry requirements Not applicable.
Mode of delivery Face-to face.
Work placements Not applicable
Learning outcomes This course unit gives students an overall perspective in molecular biology and its applications, with a strong practical component, in order to acquire skills in scientific research, molecular diagnosis, and forensic science.
By the end of this course the students must be able:
1–To describe the chemical and biological bases underlying the techniques of molecular biology and their practical application;
2–To describe the techniques for isolation and manipulation of nucleic acids;
3–To describe the methods for analysis of gene expression and its applications;
4–To execute the main techniques of molecular biology: DNA isolation, PCR, agarose gel electrophoresis, restriction and cloning;
5-To integrate the advanced concepts of molecular biology and to apply them to forensic sciences and to molecular diagnostic, planning experiments with critical spirit.
Syllabus Theoretical 1.Molecular biology: History 2.Molecular biology molecules: nucleic acids and proteins; -Structure -Prokaryotic and eukaryotic genes 3. Isolation of DNA and RNA 4. Nucleic acid manipulation-Electrophoresis -PCR -Nucleases and modification enzymes- Cloning vectors and recombinant molecules -DNA sequencing Identification / cloning of genes from DNA banks 5. Analysis of gene and protein expression - RT-PCR and real-time PCR - Hybridization, blotting and in situ hybridization - Subtractive hybridizations and DNA microarrays -Next Generation RNASequencing (RNAseq) -Transfection with a reporter gene -Western blot and immunocytochemistry 6. Applications of Molecular Biology -Genetic markers, DNA fingerprint and forensic sciences -Analysis of nucleic acids and molecular diagnostics - Small RNAs (sRNA) and plant breeding - Transgenic organisms - Genome release: CRISPR-Cas technology.

Pratical 1.Isolation of total RNA from rat cerebral cortex 2.Synthesis of cDNA. 3. PCR and electrophoresis 4.Cloning in pNZY28A 5.Transformation of E. coli. 6. Preparation of DNAp. 7.Restriction of pDNA and analysis. 7. Analysis of clone sequencing. 8.Bioinformatics: - Genetic data banks; - Sequence identification (NCBI); -notation of genomes; sequence comparison (Genbank data); -separation of BLAST sequences; - search of mutations and cut sites for restriction enzymes; - primers' design. 9. Problem solving on theoretical contents.
Main Bibliography 1.Required
- Lodish H, Berk A, Zipursky SL, et al. “Molecular Cell Biology”. 4th or 5th Ed. New York: W. H. Freeman, 2000 or 2004(http://bcs.whfreeman.com/lodish5e/)
-Alberts B., Bray D., Hopkin K. et al, “Fundamentos de Biologia Celular”. 3th Ed. Artmed, 2011.

2.Recommended
- C. Arraiano e A. Fialho, “O mundo do RNA”, 1ªEd, Lidel, 2007
- Carlos Azevedo,“Biologia Celular e Molecular”, , 4ªEd, Lidel, 2005
-“Cell Biology”,3th Ed ; A Laboratory Handbook ; Edited by: Julio E. Celis, Copyright, Elsevier Inc., 2006
- PubMed: http://www.ncbi.nlm.nih.gov/pubmed/
-Blast: http://blast.ncbi.nlm.nih.gov/Blast.cgi
Teaching Methodologies and Assessment Criteria In theoretical classes, oral exposure of the subjects will be done using data-show, with an intense student-teacher
interaction. In order to ensure participation and motivation of the students, the debate between students and the
teacher will be encouraged. In the laboratory, students will have the opportunity to perform the basic techniques of
molecular biology, which enable them to understand all the applications. They also will solve practical exercises related
to the subject taught in theoretical classes.
Evaluation:
• Theoretical component (80% = 16 points).Corresponds to the results obtained in two written tests, 8 points each
(excluding: average <6 points)
• Practical component (20% = 4 points). Corresponds to the laboratory performance and to a written report
about the laboratory work .
Regular attendance: 70% in theoretical classes, 90% in practical classes.
Language Portuguese. Tutorial support is available in English.
Last updated on: 2020-01-13

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