Nucleic acid extraction method

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/10 (2006.01)

Patent

CA 2541911

The invention relates to a method for the extraction of nucleic acids from microorganisms and to the use thereof in a method for the analysis of a microbial population in a given environment, e.g. a population of microorganisms taken from the air. In particular, the invention relates to an extraction method in which the following three lysis steps are performed successively in any order, namely: (i) chemical lysis of the microorganisms, consisting in bringing the microorganisms into contact with a lysis solution containing a detergent; (ii) thermal shock lysis comprising the incubation of the extract at a temperature lower than 0 ·C, immediately followed by the incubation of the extract at a temperature of at least 95 ·C and preferably 100 ·C; and mechanical lysis in which the extract is stirred in the presence of balls.

La présente invention porte sur une méthode d'extraction d'acides nucléiques de micro-organismes, et son utilisation dans un procédé d'analyse d'une population microbienne d'un environnement donné, notamment d'une population de micro-organismes prélevés dans l'air. En particulier, l'invention porte sur une méthode d'extraction comprenant la mise en .oelig.uvre successive des trois étapes de lyse suivantes dans un ordre quelconque : une lyse chimique des micro-organismes comprenant la mise au contact des micro-organismes avec une solution de lyse contenant un détergent ; une lyse par choc thermique comprenant l'incubation de l'extrait à une température inférieure à 0~ C, suivie immédiatement de l'incubation de l'extrait à une température d'au moins 95~ C, de préférence environ 100~ C ; et ; une lyse mécanique comprenant l'agitation de l'extrait en présence de billes.

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