Contents

The content of this module is as follows:

1) Introduction to basic molecular biology techniques.

1.1. Principles of gene cloning and DNA analysis.

- Amplification, labeling and detection of nucleic acids.
- Type of vectors, DNA molecular cloning strategies and gene libraries of DNA.
- DNA-directed mutagenesis.

1.2. Applications of gene cloning and DNA analysis for the study of gene expression.

- Techniques for studying gene expression based on DNA microarrays and massive sequencing of DNA (RT-PCR, Run On, microarrays and DNA footprinting, promoter analysis using reporter genes, mRNAs massive sequencing December, ChIP-Seq, GRO-Seq, etc.)..

2) Characteristics of commonly used model organisms. 

3) Techniques for the study of epigenetic mechanisms that regulate chromatin structure and its role in the replication, transcription and repair of eukaryotic DNA.

- Determination of heterochromatin and euchromatin regions (microscopic / sensitivity to nucleases / density gradients / chromatin precipitation curves, etc.).

- Modifications of histones (histone code). ChIP, ChIP-chip, ChIP-seq and others.

- DNA methylation. Identification of methylation in a sequence. Characterization of the degree of methylation at CpG islands. Building the methyloma.

- Methods for characterizing high and low resolution nucleosome positioning and identification of sites of hypersensitivity to nucleases (End labeling, in vivo footprinting, LM-PCR, etc.).

- Methodology for the study of chromatin remodeling complexes.

- Techniques for studying the 3D organization of the genome (Hi-C).

- Applications of epigenetics in the analysis of genomic imprinting in plants.

4) Modification of genomes and gene silencing (RNA antisense techniques, KO, ribozymes, GM, using adjustable promoters, etc.).

5) Protein Expression. Characteristics of various protein expression systems (types of vectors, promoters, organizations,etc.). Study of the intracellular localization of proteins.

6) Detection of protein-protein interaction (double hybrid protein chips, FRET, etc.) and Interactomics.

7) Applications of recombinant DNA technology in industry and medicine (diagnosis, antibody engineering, metabolic, etc.).

8) Presentation and defense of a bibliographic work.

9) Case reports.

10) Practical sessions: 24h of laboratory work divided in 8 sessions of variable length (2-4h/session). Genome editing using CRISPR-Cas9, PCR, RNA-seq analysis, chromatin immunoprecipitation, qPCR, sequence databases and analysis tools.

11) Experts seminaries.