C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/10 (2006.01) C07H 21/04 (2006.01) G01N 27/447 (2006.01)
Patent
CA 2407270
The invention concerns a method for indirectly extracting DNA target with a size greater than 300 kB from non-cultivable organisms contained in an environmental sample which consists in: grinding said sample in liquid medium; recuperating the resulting ground material; separating the organisms from the rest of the ground material by sedimentation on a cushion with density higher than 1; recuperating the organisms isolated at the surface of the cushion and incorporating it in an agarose block; extracting the DNA by lysis in the agarose block; placing the block in an agarose gel which is subjected to at least pulsed field electrophoresis; finally, recuperating the extracted DNA strands.
Procédé d'extraction indirecte de d'ADN cible de taille supérieure à 300 kB d'organismes non cultivables contenus dans un échantillon environnemental selon lequel: on broie en milieu liquide ledit échantillon; on récupère le broyât obtenu; on sépare les organismes du reste du broyât par sédimentation sur coussin de densité supérieure à I; on récupère les organismes isolés à la surface du coussin que l'on inclut dans un bloc d'agarose; on extrait l'ADN par lyse dans le bloc d'agarose; on positionne le bloc dans un gel d'agarose que l'on soumet à au moins une électrophorèse en champs alternés, enfin, on récupère les brins d'ADN extraits.
Nalin Renaud
Robe Patrick
Tran Van Van
Brouillette & Associes/partners
Libragen
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