|
|
| |
|
Molecular biology
Equipment
- Bacterial culture: thermo-regulated rotating incubators, centrifuges
- DNA/RNA analysis: apparatus for DNA/RNA/protein purification; agarose and acrylamide electrophoresis;
digital picture acquisition, quantitative analysis software
- PCR amplification: PCR apparatus, PCR hood
- Protein analysis: 1D and mini-2D Western Blot analysis, DotBlot, ELISA, FACS
- Miscellaneous: Stratalinker, sonication disruptor, mixer mill disruptor
- Analysis: Spectrometer, Fluorometer, Luminometer
|
Experiments performed by the laboratory
To observe : Quantify gene and protein expression.
- RT/PCR analysis (RNA expression by semi-quantitative and quantitative methods, sequence specific amplification, sensitive detection, optimization of methodology
- WB analysis: semi-quantitative methods, optimization of methodology
To modify : Modify gene and protein expression.
- cDNA isolation
- Mutagenesis
- Constitutive or inducible expression
- Cellular cloning (lipofectant transfection of siRNA and shRNA)
To characterize : Use functional assays to observe molecular action.
- Molecular assays : immunoprecipitation, gel shifts and Dot Blot
- Cellular assays : GENE reporter
|
Aim of these experiments in drug development
To observe, to modify and to characterize gene and protein expression.
Gene expression and target validation = to observe
- Evaluation of expression in a panel of cultured cell lines and grafted or primary tumors
- Evaluation of gene or protein expression after in vitro or in vivo treatment
- Detection of organ invasion by tumor cells
- Evaluation of antibody specificity
- Biomarker follow-up
Structure-function studies = to modify
- Transient or stable over-expression of wild-type or mutated proteins in constitutive or inducible systems
- Decrease of expression by RNA interference: RNAse III siRNA, synthetic siRNA and plasmidic shRNA(constitutive or inducible)
Interaction studies = to characterize
- Immunoprecipitation (Tag or specific antibodies)
- Gel-shift, filter binding assay, Dot Blot, GENE reporter
|
|
|
Home
Back
Top