Methods and compositions for the preparation and use of...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) G01N 33/00 (2006.01)

Patent

CA 2532467

This invention relates to methods for the detection of one or more mRNA transcripts in paraffin-embedded tissue by "mRNA liberation in fixed-treated tissue or 'MLIFTT"'. This method includes treating the tissue with ammonia- ethanol and sodium borohydride combined with pressure cooking of the tissue. The chemical treatments reduce the tissue autofluorescence and the physical treatments overcome the interference created by the fixation-induced chemical bonds. The methods of the present invention can be utilized to identify a plurality of mRNA transcripts in a microarray format.

L'invention concerne des procédés pour la détection d'un ou de plusieurs produits de transcription d'ARNm dans un tissu noyé dans de la paraffine, par libération d'ARNm dans un tissu traité fixé, technique appelée <= MLIFTT >=. Ce procédé consiste à traiter le tissu avec de l'ammoniac/éthanol et du borhydrure de sodium et à réaliser une cuisson sous pression du tissu. Les traitements chimiques réduisent l'autofluorescence du tissu et les traitements physiques permettent de maîtriser l'interférence créée par les liaisons chimiques induites par fixation. Les procédés selon l'invention peuvent être mis en oeuvre pour identifier une pluralité de produits de transcription d'ARNm dans un format de microréseau.

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