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Biochemistry
Applied Molecular Biology

Applied Molecular Biology

Code	17228
Year	1
Semester	S1
ECTS Credits	6
Workload	TP(60H)
Scientific area	Biochemistry
Entry requirements	Not aplicable
Learning outcomes	THEORETICAL 1. Molecules of Molecular Biology: nucleic acids, proteins. Prokaryotic and eukaryotic genomes. 2. Isolation of DNA and RNA. 3. Nucleic acid manipulation: Electrophoresis; PCR; Nucleases and modification enzymes; Cloning vectors and recombinant molecules; DNA sequencing; Identification of gene banks. 4. Gene and protein expression: RT-PCR and real-time PCR; Hybridization, blotting, in situ hybridization; Subtractive hybridization and DNA microarrays; Next-generation RNA sequencing, Western blot; immunocytochemistry. 5. Applications of Molecular Biology: Genetic markers; DNA fingerprinting and forensic science; Nucleic acid analysis and molecular diagnostics; Small RNAs; Genome editing - CRISPR-Cas technology. PRACTICE (laboratory classes): isolation of total RNA from rat liver; cDNA synthesis; PCR/electrophoresis; Cloning into a vector suitable for PCR products; E. coli transformation; pDNA isolation; Restriction/digestion product analysis. Sequencing.
Syllabus	THEORETICAL 1. Molecules of Molecular Biology: nucleic acids, proteins. Prokaryotic and eukaryotic genome 2. Isolation of DNA and RNA 3. Manipulation of nucleic acids: Electrophoresis; PCR; Nucleases and modification enzymes; Cloning vectors and recombinant molecules; DNA sequencing; Identification of gene banks 4. Expression of genes and proteins: RT-PCR and real-time PCR; Hybridization, blotting, in situ hybridization; Subtractive hybridizations and DNA microarrays; Next-generation RNA sequencing, Western blot; immunocytochemistry 5. Applications of Molecular Biology: Genetic markers; DNA fingerprinting and forensic science; Nucleic acid analysis and molecular diagnostics; Small RNAs; Genome editing - CRISPR-Cas technology. PRACTICE (laboratory classes): isolation of total RNA from rat liver; cDNA synthesis; PCR/electrophoresis; Cloning of vector suitable for PCR products; E. coli transformation; pDNA isolation; Restriction/digestion product analysis. Sequencing
Main Bibliography	1. Main Bibliography - Benjamin A. Pierce. "Genetics - A Conceptual Approach," 7th Edition, 2020. W. H. Freeman Publishers - Lodish H, Berk A, Zipursky SL, et al. "Molecular Cell Biology." 4th or 5th Ed. New York: W. H. Freeman, 2021 - Alberts B., Bray D., Hopkin K. et al., "Fundamentals of Cell Biology." 4th Ed. Artmed, 2017. 2. Complementary Bibliography - José Luque, Angel Herraez, "Molecular Biology and Genetic Engineering" - Concepts, Techniques, and Applications in Health Sciences. Elsevier, 2006 - Carlos Azevedo, “Cellular and Molecular Biology,” 4th Ed., Lidel, 2005 - “Cell Biology,” 3rd Ed.; A Laboratory Manual; 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
Language	Portuguese. Tutorial support is available in English.