Method for the isolation of mrna from formalin fixed,...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2626533

The invention provides a reliable method for recovering RNA from a formalin- fixed, paraffin-embedded biological tissue sample comprising: 5 a) deparaffinizing the sample, b) contacting said sample with a solution comprising an effective concentration of proteinase K and heating the sample in said solution to a temperature in the range of about 30 to about 60 ~C, for a time period of about 12 to 20 hours, c) adding an effective concentration of proteinase K to the solution obtained from 10 b) for a time period of about 5 to about 30 minutes at a temperature range of about 40 to about 75 ~C, and d) recovering said RNA from said solution. The additional step c) of digestion improves significantly the recovery of RNA.

L'invention concerne un procédé fiable servant à récupérer l'ARN à partir d'un échantillon de tissu biologique enrobé de paraffine et fixé avec de la formaline, consistant : a) à déparaffiner l'échantillon, b) à mettre en contact ledit échantillon avec une solution comprenant une concentration efficace de protéinase K et à chauffer l'échantillon dans ladite solution à une température comprise dans la plage d'environ 30 à environ 60°C, pendant une durée d'environ 12 à 20 heures, c) à ajouter une concentration efficace de protéinase K à la solution obtenue à partir de b) pendant une durée d'environ 5 à environ 30 minutes à une température comprise dans la plage d'environ 40 à environ 75°C et d) à récupérer ledit ARN à partir de ladite solution. L'étape supplémentaire c) de digestion améliore significativement la récupération de l'ARN.

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