Theoretical part (32 hours)
Introduction to nucleic acids and proteins handling:
Methods for DNA, RNA and proteins isolation from biological samples and related analysis. Comparison between protein samples: differential-in-gel electrophoresis (DIGE).
Methods for gene expression analysis:
Semi-quantitative, competitive and quantitative PCR. DNA microarray technology: cDNA and oligonucleotide microarrays,SAGE, Subctractive Library.
Methods for protein-protein interaction analysis:
Display in vitro: Protein Arrays, Far-Western, Pull-down, mRNA display, Ribosome display.
Display in vivo: Co-Immunoprecipitation, Tandem Affinity Purification (TAP) system, Phage Display, Bimolecular Fluorescent Complementation (BiFC).
Methods for DNA/RNA-protein interaction analysis:
Chromatin Immunoprecipitation assay (ChIP and ChIP-on-chip). Electrophoretic Mobility Shift Assay (EMSA). Southwestern. Yeast three-hybrid system.
Methods for epigenetic modifications analysis :
Methylation-sensitive restriction enzymes-based techniques: Methylation-Sensitive Amplification Polymorphism (MSAP). Bisulfite conversion and subsequent methods of analysis: Bisulfite (non methylation)-specific PCR and Methylation-specific PCR (MSP). Methods for histone modifications analysis.
Mutagenesis techniques:
Site-directed and random mutagenesis: primer-extension mutagenesis and PCR-based mutagenesis. Mutagenesis by homologous recombination: DNA Shuffling.

Genome editing techniques:
Zinc finger nuclease (ZFN), Transcription activator–like effector-based nuclease (TALEN) e CRISPR-Cas9 System.
Diagnostic and forensic application of PCR.
DNA Sequencing. Human genome project.
Next Generation Sequencing: second and third generation sequencing platforms.

Practical part (16 hours)

Extraction of total RNA from Arabidopsis leaves, RNA quantification and analysis on agarose gel
Genomic DNA extraction from Arabidopsis leaves, DNA quantification, and analysis on agarose gel.
Extraction of water-soluble proteins from Arabidopsis leaves and quantification of protein extract using the Bradford method. In batch purification of GST (Glutathione S-Transferase) protein using GSH conjugated magnetic beads. Analysis of purified protein.

Brown T.A. Gene cloning and DNA analysis: an introduction. 7th ed., 2016, Wiley-Blackwell.
Watson J.D., Caudy A.A., Myers R.M., Witkowski J.A. Recombinant DNA, genes and genomes – a short course. 3rd ed., 2007, W.H. Freeman & Co.
Lesk A.M. Introduction to Genomics. 3rd ed., 2017, Oxford University press.

Slides are available in the teaching platform. Handouts are provided by the teacher for practical laboratory activities.
Non-attending students are encouraged to contact the teacher for information about the program, teaching materials, and the examination mode.