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, Ajouter 400 µL de tampon AMMCGE (1.1 : 1) (Chlorure de guanidium 7.8M / Ethanol). Mélanger soigneusement par pipetage

, Remarque : à cette étape, il est possible de laisser l'échantillon à 4°C pendant quelques heures

, Appliquer les 750 ?L du mélange dans la plaque de fixation (Plaque Acroprep 96 Filter Plate PALL (ref PN 5051)). Centrifuger la plaque 2 min à 6000 rpm à température ambiante

, Remarque : certains échantillons contiennent des particules en suspension qui peuvent colmater les membranes

, 1° lavage: déposer 600 ?L de tampon AMMLav/E (Ethanol et sel d'acétate de potassium) dans chaque puits de la plaque

, Eliminer le liquide qui passe. Remarques : -les traces résiduelles d'éthanol sur la membrane interfèrent avec le tampon d'élution et peuvent faire chuter le rendement de récupération de l'ADN, 2° lavage: déposer 600 ?L de tampon AMMLav/E (Ethanol et sels d'acétate de potassium) dans chaque puits de la plaque

, Les membranes peuvent céder à la force centrifuge et se percer d'un trou (de petit diamètre) en leur centre. Le rendement de récupération de l'ADN n'est pas altéré

E. De-l'adn-1, Déposer sur la colonne 100 ?L d'eau ultrapure (ou de TE1X) préchauffé vers 50°C. Attendre 5 min à température ambiante

, Centrifuger 2 min à 6000 rpm à température ambiante. Récupérer le liquide qui passe dans une plaque de réception

, plaque 96 de stockage en faible volume en polypropylène, plaque 96 puits profonds ou tubes Micronic courts sur portoir 96. Faire attention à la résistance mécanique de la plaque de réception lors de la centrifugation à 6200 g, PREPARATION DES SOLUTIONS Tampon SDS : Pour 100 mL mélanger : 20 mL de Tris 1M (pH = 8), vol.10

, Pour 100 mL mélanger : 60 ml d'acétate de potassium 5M, 11.5 mL d'acide acétique glacial (37%)


, Produit Sigma ref G-3272) : Dissoudre 37.25 g de chlorure de guanidium dans l'eau jusqu'à un volume final de 50 mL de solution (faire le mélange dans un tube Sarstedt ou Falcon de 50 mL). Vérifier, si possible, Préparation de la solution de chlorure de guanidium 7.8 M

, Tris HCl ph 8 22,5 mM, EDTA 0,1 mM) mélanger : 2,25 mL de Tris 1M pH = 8, Pour 100 mL d'une solution aqueuse d'AMMlav (Acétate de potassium 160 mM