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, Annexe 5 : protocole d'extraction d'ARN

, Ajouter 450µL de tampon RLC + 4,5µL de ?-mercaptoéthanol et vortexer

, Ajouter 0,5V d'éthanol ARN free et agiter par retournement doucement

, Laver la membrane avec 350µL de RW1, centrifugation 30 secondes à 10 000rpm

T. Dnase, Ajouter 80µL d'un mélange DNase + tampon

, Ajouter 350µL de RW1, centrifugation 30 secondes à 10 000rpm

, Vider la partie liquide

, Ajouter 500µL de RPE et centrifuger 30 secondes à 10 000rpm

, Vider la partie liquide et remettre 350µL de RPE + centrifugation 30 secondes à, vol.10, p.0

, Enlever le tube de récupération, et le remplacer par un eppendorf ARN free

, Centrifuger 1 minute pour récupérer l'ARN dans l'eppendorf

, Ajouter 200µL de LiCl 4M (permet la dégradation de potentielle RNases), retourner doucement 20) Stocker à-20°C