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L'invention concerne un procédé pour l'analyse quantitative in vitro et/ou ex vivo du nombre de molécules d'acides nucléiques codant pour des gènes différents exprimés au niveau de cellules d'organismes vivants, sur un petit nombre de cellules, de préférence sur une seule cellule, dans lequel:- on effectue la transcription inverse et une première réaction d'amplification en chaîne par polymérase (par RT-ACP) de l'ARNm d'un faible nombre de cellules lysées,- on sépare les divers produits de ladite première réaction d'amplification en chaîne par polymérase pour individualiser les gènes, et- on conduit une deuxième réaction d'amplification en chaîne par polymérase sur les gènes individuels ainsi séparés, tandis que:- ladite première réaction d'amplification en chaîne par polymérase est effectuée au moyen d'une association d'amorces 5' et d'amorces 3', lesdites amorces 3' étant identiques aux amorces 3' utilisées pour la transcription inverse,- on utilise, comme référence pour le calcul du nombre de copies de gènes générées par amplification par polymérase, l'ADN cellulaire individuel codant pour le gène considéré,- on contrôle l'efficacité de la transcription inverse, au moyen d'une amplification en chaîne par polymérase des ADNc de l'un des gènes concernés, parallèlement à la même amplification des molécules d'ARNm du même gène, sur des échantillons cellulaires contenant le même nombre de molécules, et- on évalue les résultats de ladite deuxième réaction d'amplification.
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