1. TECHNICAL SKILLS
- Human blood handling: plasma separation from human whole blood; isolation of peripheral blood
mononuclear cells by Ficoll-Paque gradient; DNA isolation from human blood samples.
- Mutation discovery and polymorphism genotyping: Polymerase Chain Reaction (PCR); Restriction
Fragment Length Polymorphisms (RFLP) analysis; TaqMan Allelic Discrimination Real-Time PCR; horizontal
gel electrophoresis; Southern Blotting; denaturing high-performance liquid chromatography (dHPLC);
Sequencing (ABI 3130 XL DNA sequencer).
- RNA/Protein expression: RNA/Protein extraction and quantification (from tissue/cell cultures); relative
quantitative Real-Time PCR; Enzyme Linked Immunosorbent Assay (sandwich); Western Blotting and Dot
Blotting.
- Molecular Biology: Restriction digest, plasmid construction, cloning and transformation in Escherichia coli
and mutagenesis. Cultures of Saccharomyces cerevisiae (wild-type and mutant strains); centrifugal
elutriation.
- Cell cultures: Manipulation of mammalian cell lines; transfection and clonal selection; vitality assays;
immunocytochemistry.
- Animal models: Breeding, weaning and genotyping of wild-type and transgenic mouse models for
Alzheimer’s disease (APP23,CRND8) and Tauopathies (P301L); intraperitoneal injection of candidate drugs;
intracardiac perfusion; behavioral tests (Morris Water Maze, Novel Object Recognition Test) ̧
immunohistochemistry.