Yeast specific techniques

∙ Yeast cells genetic engineering by homologous recombination

∙ Robotic handling of yeast deletion and two-hybrid strains collections with the Biomek 3000 system

∙ RNA metabolic labeling with 5,6-³H-uracil

∙ DNA/RNA extraction methods

∙ Native/Denaturing protein extraction methods

Biochemistry

∙ Protein overexpression in bacteria and yeast cells

∙ Fast performance liquid chromatography

∙ Immunoprecipitation and affinity purification techniques (GST, Protein A, His tags) under native conditions

∙ In vitro kinase assays with purified substrates or peptides.

∙ LacZ/luciferase reporter assays

∙ Denaturing protein extraction for optimal conservation of phosphorylation by the TCA-Urea method

∙ Protein phosphorylation analysis by SDS-PAGE migration shift, by Phos-Tag-induced migration shift and by direct staining with fluorescent dyes (ProQ Diamond staining)

∙ Handling of radioactive isotopes: ³H, ¹⁴C and ³²P.

Biology

∙ Mammalian cells culture and handling (HeLa, 293, T47D, SkBr3, MCF7)

∙ Mammalian cells transfection by the calcium/phosphate method, by lipofection or with the pLNCX retroviral vector system

∙ Viral amplification in fertilized chicken eggs allantoic cavity

∙ Plasmids engineering and mutagenesis

∙ Wide-field and fluorescence microscopy on fixed and live cells (yeast/mammalian cells) and fixed tissues

∙ Confocal fluorescence microscopy on fixed cells (Leica SP2)

∙ Chromatin immunoprecipitation (ChIP)

∙ SYBR green quantitative RT-PCR (Roche LightCycler 480)

Computer sciences

∙ Experience on Windows and Linux-based systems

∙ Visual Basic programming (Standalone applications, Excel development)

∙ PHP 5 - MySQL web development