Fluorescent probes for chromosomal painting

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2345381

The invention concerns fluorescent probes used in multicolour in situ fluorescent hybridisation methods, and principally chromosomal painting. The probes designed for marking a chromosome, are such that they consist of a set of DNA segments more represented in certain chromosomal bands and are obtained by IRS-PCR amplification from said chromosomes using PCR primers specific of the repeated and dispersed DNA sequences Alu and LINE. The invention further concerns methods for producing said probes, multicolour FISH methods capable of using said probes and diagnosis kits comprising them. The invention also concerns combinations of fluorophores and optical filters.

La présente invention concerne des sondes fluorescentes utilisables dans les méthodes d'hybridation fluorescente in situ multicouleur, et principalement la peinture chromosomique. Les sondes destinées au marquage d'un chromosome, sont telles qu'elles sont composées d'un ensemble de segments d'ADN davantage représentés dns certaines bandes chromosomiques et sont telles qu'elles sont obtenues par amplification IRS-PCR à partir desdits chromosomes en utilisant des amorces PCR spécifiques des séquences d'ADN répétées et dispersées Alu et LINE. L'invention comprend, en outre, des procédés de production desdites sondes, des procédés FISH multicouleur pouvant mettre en oeuvre lesdites sondes ainsi que des kits de diagnostic les comprenant. Enfin, l'invention comprend des combinaisons de fluorophores et de filtres optiques.

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