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Development of imaging devices and methods for high sensitivity quantification of dna by chemiluminescence

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In our project we propose to improve the current limitations of quantitative and high sensitive imaging of nucleic acids on solid supports by chemiluminescence. First, using a back illuminated, cooled CCD-camera (charge coupled device), measurements will de- termine the required physico-chemical conditions for quantitative imaging of chemiluminescent probes immobilised on solid supports (membranes) at low light levels. Simultaneously, to reach the ex- treme sensitivity required for certain applications, a second imager, based on an intensified PSD (position sensitive detector), will be developed. In addition, the already patented technology STIC (self triggered intensified CCD), which was developed by the host group, will be implemented in this device. Those modifications are expected to result in the following specifications, to image and quantify, after hybridisation with a non radioactive probe, DNA immobilised on membranes:
? Surface analysed : 20x20 cm2
? Sensitivity : < 3 fg/mm2
? Dynamic range : 5000
? Spatial resolution : < 0.5 mm
Finally, this apparatus will be used in a differential screening experiment on c-jun targets, to test a model of neurobiology. Moreover, molecular genetics in the process of development of Drosophila melanogaster and molecular modifications in the cen- tral nervous system of vertebrates, induced by cocaine and am- phetamines, are planed to be studied.

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RGI - Research grants (individual fellowships)

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CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
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Rue Georges Clémenceau Université de Paris-Sud 15
91406 ORSAY
Frankreich
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